METHODS AND KITS FOR DETECTING SEVERE ACUTE RESPIRATORY SYNDROME CORONAVIRUS 2 FIELD The disclosure relates to a kit for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The disclosure also relates to a method for detecting the presence of SARS-CoV-2 in a biological sample using the kit. BACKGROUND Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an enveloped, positive-sense, single-stranded RNA betacoronavirus that causes a pandemic of acute respiratory disease, denoted as coronavirus disease 2019 (COVID-19). SARS-CoV-2 has a large viral RNA (vRNA) genome which contains 6– 11 open reading frames (ORFs) within the 5’ and 3’ flanking untranslated regions (UTRs). The two main transcriptional units, ORF1a and ORF1ab, encode replicase polyprotein 1a (PP1a) and polyprotein 1ab (PP1ab), respectively. The largest polyprotein PP1ab embeds 16 non-structural proteins (i.e., Nsp1 to Nsp16) that are involved in transcription and virus replication. The remaining ORFs encode four major structural proteins, namely, surface spike (S), transmembrane (M), nucleocapsid (N), and envelope (E). Major symptoms of COVID-19 include respiratory symptoms such as fever above 38°C, cough, shortness of breath, and difficulty in breathing. Symptoms such as loss of smell and taste, diarrhea, headache, chills, loss of appetite, general malaise, and impaired consciousness may be observed. However, more than 20% of patients with COVID-19 are asymptomatic carriers. Therefore, how to detect these potential spreaders is a challenge for alleviating and even stopping the COVID-19 pandemic. At present, several molecular techniques are applied to detect SARS-CoV-2, including reverse transcription-polymerase chain reaction (RT-PCR), real-time quantitative RT-PCR (real-time qRT-PCR), RT-loop-mediated isothermal amplification (RT-LAMP), etc. As reported in Huang, W. E. et al. (2020), Microb. Biotechnol., 13:950-961, Huang, W. E. et al. designed four primer sets, namely N1, N15, S17, and O117 (each including six primers), based on the regions of ORF1ab, S gene, and N gene of the SARS- CoV-2 genome, and such primer sets can be used in RT-LAMP assay for the detection of SARS-CoV-2 in clinical samples. In spite of the aforesaid, researchers in this field are still endeavoring to look for more satisfactory nucleic acids for the rapid and accurate detection of SARS-CoV-2. SUMMARY Therefore, in a first aspect, the present disclosure provides a method for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV- 2) in a biological sample, which can alleviate at least one of the drawbacks of the prior art. The method includes: subjecting nucleic acids in the biological sample to a nucleic acid amplification reaction with a reaction mixture that includes at least one primer set for amplifying a target nucleic acid of the SARS- CoV-2; and detecting presence or absence of an amplification product obtainable from the nucleic acid amplification reaction, wherein the presence of the amplification product is indicative of presence of the SARS-CoV-2 in the biological sample. The at least one primer set is selected from the group consisting of: (a)a first primer set for amplifying a region of nonstructural protein 2 (nsp2) gene of the SARS- CoV-2, the first primer set including a first forward outer primer having a nucleotide sequence of SEQ ID NO: 1 and a first backward outer primer having a nucleotide sequence of SEQ ID NO: 2; (b) a second primer set for amplifying a region of nsp2 gene of the SARS-CoV-2, the second primer set including a second forward outer primer having a nucleotide sequence of SEQ ID NO: 7 and a second backward outer primer having a nucleotide sequence of SEQ ID NO: 8; (c)a third primer set for amplifying a region of nonstructural protein 4 (nsp4) gene of the SARS- CoV-2, the third primer set including a third forward outer primer having a nucleotide sequence of SEQ ID NO: 13 and a third backward outer primer having a nucleotide sequence of SEQ ID NO: 14; (d) a fourth primer set for amplifying a region of nsp4 gene of the SARS-CoV-2, the fourth primer set including a fourth forward outer primer having a nucleotide sequence of SEQ ID NO: 19 and a fourth backward outer primer having a nucleotide sequence of SEQ ID NO: 20; and (e)a fifth primer set for amplifying a region of spike (S) gene of the SARS-CoV-2, the fifth primer set including a fifth forward outer primer having a nucleotide sequence of SEQ ID NO: 25 and a fifth backward outer primer having a nucleotide sequence of SEQ ID NO: 26. In a second aspect, the present disclosure provides a kit for detecting SARS-CoV-2, which can alleviate at least one of the drawbacks of the prior art. The kit includes at least one of the abovementioned primer sets for amplifying a target nucleic acid of the SARS-CoV-2. BRIEF DESCRIPTION OF THE DRAWINGS The above and other objects, features and advantages of the present disclosure will become apparent with reference to the following detailed description and the exemplary embodiments taken in conjunction with the accompanying drawings, in which: Fig. 1 shows the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with loop-mediated isothermal amplification (LAMP) using the primer set nsp2-1, in which: the upper panel is a digital image of the result of the gel electrophoresis assay of LAMP reaction products, where Lane M=100-3000 bp size marker, and Lane NTC=no template control (water); and the lower panel is a digital image of the result of the colorimetric assay using phenol red; Fig. 2 shows the detection of SARS-CoV-2 with LAMP using the primer set nsp2-2, in which: the upper panel is a digital image of the result of the gel electrophoresis assay of LAMP reaction products, where Lane M=100-3000 bp size marker, and Lane NTC=no template control (water); and the lower panel is a digital image of the result of the colorimetric assay using phenol red; Fig. 3 shows the detection of SARS-CoV-2 with LAMP using the primer set nsp4-1, in which: the upper panel is a digital image of the result of the gel electrophoresis assay of LAMP reaction products, where Lane M=100-3000 bp size marker, and Lane NTC=no template control (water); and the lower panel is a digital image of the result of the colorimetric assay using phenol red; Fig. 4 shows the detection of SARS-CoV-2 with LAMP using the primer set nsp4-2, in which: the upper panel is a digital image of the result of the gel electrophoresis assay of LAMP reaction products, where Lane M=100-3000 bp size marker, and Lane NTC=no template control (water); and the lower panel is a digital image of the result of the colorimetric assay using phenol red; Fig. 5 shows the detection of SARS-CoV-2 with LAMP using the primer set S-1, in which: the upper panel is a digital image of the result of the gel electrophoresis assay of LAMP reaction products, where Lane M=100-3000 bp size marker, and Lane NTC=no template control (water); and the lower panel is a digital image of the result of the colorimetric assay using phenol red; Fig. 6 shows the detection of SARS-CoV-2 with LAMP using the primer set S-2, in which: the upper panel is a digital image of the result of the gel electrophoresis assay of LAMP reaction products, where Lane M=100-3000 bp size marker, and Lane NTC=no template control (water); and the lower panel is a digital image of the result of the colorimetric assay using phenol red; Fig. 7 shows the detection of SARS-CoV-2 with reverse transcription loop-mediated isothermal amplification (RT-LAMP) using a combination of the primer sets nsp2-2 and nsp4-2, in which: the upper panel is a digital image of the result of the gel electrophoresis assay of RT-LAMP reaction products, where Lane M=100-3000 bp size marker, and Lane NTC=no template control (water); and the lower panel is a digital image of the result of the colorimetric assay using phenol red; Fig. 8 shows the detection of SARS-CoV-2 with RT-LAMP using the combination of the primer sets nsp2-2 and nsp4-2, in which: the upper panel is a digital image of the result of the gel electrophoresis assay of RT-LAMP reaction products, where Lane M=100-3000 bp size marker, and Lane NTC=no template control (water); and the lower panel is a digital image of the result of the colorimetric assay using phenol red; and Fig. 9 shows the detection of SARS-CoV-2 with RT-LAMP using the combination of the primer sets nsp2-2 and nsp4-2, in which: the upper panel is a digital image of the result of the gel electrophoresis assay of RT-LAMP reaction products, where Lane M=100-3000 bp size marker, and Lane NTC=no template control (water); and the lower panel is a digital image of the result of the colorimetric assay using phenol red. DETAILED DESCRIPTION It is to be understood that, if any prior art publication is referred to herein, such reference does not constitute an admission that the publication forms a part of the common general knowledge in the art, in&KLQD or any other country. For the purpose of this specification, it will be clearly understood that the word “comprising” means “including but not limited to”, and that the word “comprises” has a corresponding meaning. Unless otherwise defined, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this disclosure belongs. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of this disclosure. Indeed, this disclosure is in no way limited to the methods and materials described. For clarity, the following definitions are used herein. As used herein, the terms “nucleic acid”, “nucleic acid sequence”, and “nucleic acid fragment” refer to a sequence of nucleotides connected by phosphodiester linkages and may be a deoxyribonucleic acid (DNA) molecule or ribonucleic acid (RNA) molecule in either single-stranded or double- stranded form. The nucleic acid may comprise naturally occurring nucleotides and known analogues thereof, as well as nucleotides that are modified in the sugar and/or phosphate moieties. This term also encompasses nucleic acids containing modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. The term “nucleic acid” as used herein is interchangeable with the terms “gene”, “cDNA”, “mRNA”, “oligo- nucleotide”, and “polynucleotide” in use. As used herein, the term “DNA fragment” refers to a DNA polymer, in the form of a separate segment or as a component of a larger DNA construct, which has been derived either from isolated DNA or synthesized chemically or enzymatically such as by methods disclosed elsewhere. Unless otherwise indicated, a nucleic acid sequence, in addition to the specific sequences described herein, also covers its complementary sequence, and the conservative analogs, related naturally occurring structural variants and/or synthetic non-naturally occurring analogs thereof. Unless denoted otherwise, whenever a nucleic acid sequence is represented, it will be understood that the nucleotides are in 5′ to 3′ order from left to right and that “A” denotes deoxyadenosine or an analog thereof, “C” denotes deoxycytidine or an analog thereof, “G” denotes deoxyguanosine or an analog thereof, and “T” denotes deoxythymidine or an analog thereof. The term “3′” refers to a region or position in a polynucleotide or oligonucleotide 3′ (i.e., downstream) from another region or position in the same polynucleotide or oligonucleotide. The term “5′” refers to a region or position in a polynucleotide or oligonucleotide 5′ (i.e., upstream) from another region or position in the same polynucleotide or oligonucleotide. The terms “3′-end” and “3′- terminus”, as used herein in reference to a nucleic acid molecule, refer to the end of the nucleic acid which contains a free hydroxyl group attached to the 3′ carbon of the terminal pentose sugar. The terms “5′-end” and “5′-terminus”, as used herein in reference to a nucleic acid molecule, refer to the end of the nucleic acid molecule which contains a free hydroxyl or phosphate group attached to the 5′ carbon of the terminal pentose sugar. As used herein, the term “primer” refers to an oligonucleotide of defined sequence that is designed to hybridize with a complementary, primer-specific portion of a target polynucleotide sequence and undergo primer extension. The primer can function as the starting point for the enzymatic polymerization of nucleotides using a polymerase. The primer should be long enough to prevent itself from annealing to sequences other than the complementary portion. Generally, the primer is between 13 to 50 nucleotides in length. Preferably, the primer is between 15 to 30 nucleotides in length. The primer used herein may also be used in nucleic acid amplification. As used herein, the term “amplification” may refer to an increase of the number of copies of a target sequence or a complementary sequence of the target sequence, such as a gene or fragment of a gene. A “copy” or “amplicon” does not necessarily mean perfect sequence complementarity or identity to template sequence. For example, copies can include nucleotide analogs such as deoxyinosine, intentional sequence alterations (such as sequence alterations introduced through a primer including a sequence that is hybridizable but not complementary to the template), and/or sequence errors that occur during amplification. The products of an amplification reaction are called amplification products. The nucleic acid amplification may be performed using any method known in the art, e.g. a method employing multiple heat cycles during the amplification, or a method performed at a constant temperature (i.e., isothermal amplification). An example of in vitro amplification requiring heat cycles is polymerase chain reaction (PCR), in which a sample (such as a biological sample from a subject) is contacted with a pair of oligonucleotide primers, under conditions that allow for hybridization of the primers to a nucleic acid molecule in the sample. The primers are extended under suitable conditions, dissociated from the template, and then re-annealed, extended, and dissociated to amplify the number of copies of the nucleic acid molecule. Other examples of in vitro amplification techniques may include, but are not limited to, PCR, quantitative PCR (qPCR), reverse transcription PCR (RT-PCR), quantitative RT- PCR (RT-qPCR), nested polymerase chain reaction, hot- start polymerase chain reaction, multiplex polymerase chain reaction, ligase chain reaction (LCR), gap ligase chain reaction (gLCR), etc. On the other hands, isothermal amplification may be performed at a constant temperature, or a major aspect of the amplification process occurs at a constant temperature, i.e., without significant changes in temperature. Thus, it is carried out substantially at about the same single temperature. In some examples, isothermal amplification is substantially isothermal, for example, may include small variations in temperature, such as changes in temperature of no more than about 1°C to 3°C during the amplification reaction. As used herein, the terms “target sequence” and “target nucleic acid” can be interchangeably used, and refer to a particular nucleic acid sequence which is to be detected and/or amplified. Preferably, target sequences include nucleic acid sequences to which the primers complex in a PCR reaction. Target sequences may also include a probe hybridizing region with which a detection probe will form a stable hybrid under desired conditions. As will be recognized by one of ordinary skill in the art, a target sequence may be single-stranded or double- stranded. In the context of the present disclosure, target sequences of interest may be within the nonstructural protein 2 (nsp2) gene, nonstructural protein 4 (nsp4) gene, or spike (S) gene of severe acute respiratory syndrome coronavirus 2 (SARS-CoV- 2). As used herein, the terms “hybridization” and “annealing” can be interchangeably used, and refer to the process in which complementary nucleic acid strands react to form a “hybrid” or “duplex” that is stabilized via hydrogen bonding between the bases of the nucleotide residues. The hydrogen bonding may occur by Watson Crick base pairing, Hoogsteen binding, or in any other sequence specific manner. As used herein, the terms “duplex” and “hybrid” can be interchangeably used, and refer to a structure formed as a result of hybridization of two complementary sequences of nucleic acids. Such duplexes can be formed by the complementary binding of two DNA segments to each other, two RNA segments to each other, or of a DNA segment to an RNA segment. Either or both members of such duplexes can contain modified nucleotides and/or nucleotide analogues as well as nucleoside analogues. As disclosed herein, such duplexes are formed as the result of binding of one or more probes to a sample sequence. The present disclosure provides a method for detecting SARS-CoV-2 in a biological sample, which includes: subjecting nucleic acids in the biological sample to a nucleic acid amplification reaction with a reaction mixture that includes at least one primer set for amplifying a target nucleic acid of the SARS- CoV-2; and detecting presence or absence of an amplification product obtainable from the nucleic acid amplification reaction, wherein the presence of the amplification product is indicative of presence of the SARS-CoV-2 in the biological sample. The at least one primer set for amplifying the target nucleic acid of the SARS-CoV-2 includes: (a) a first primer set for amplifying a region of nsp2 gene of the SARS-CoV-2, the first primer set including a first forward outer primer having a nucleotide sequence of SEQ ID NO: 1 and a first backward outer primer having a nucleotide sequence of SEQ ID NO: 2; (b) a second primer set for amplifying a region of nsp2 gene of the SARS-CoV-2, the second primer set including a second forward outer primer having a nucleotide sequence of SEQ ID NO: 7 and a second backward outer primer having a nucleotide sequence of SEQ ID NO: 8; (c) a third primer set for amplifying a region of nsp4 gene of the SARS-CoV-2, the third primer set including a third forward outer primer having a nucleotide sequence of SEQ ID NO: 13 and a third backward outer primer having a nucleotide sequence of SEQ ID NO: 14; (d) a fourth primer set for amplifying a region of nsp4 gene of the SARS-CoV-2, the fourth primer set including a fourth forward outer primer having a nucleotide sequence of SEQ ID NO: 19 and a fourth backward outer primer having a nucleotide sequence of SEQ ID NO: 20; and (e) a fifth primer set for amplifying a region of S gene of the SARS-CoV-2, the fifth primer set including a fifth forward outer primer having a nucleotide sequence of SEQ ID NO: 25 and a fifth backward outer primer having a nucleotide sequence of SEQ ID NO: 26. According to this disclosure, the nucleic acids (e.g., viral genomic RNA of the SARS-CoV-2) in the sample may be first extracted from the sample, and then subjected to the amplification reaction. The procedures and conditions for extracting nucleic acids are within the expertise and routine skills of those skilled in the art (for example, see Huang, W.E. et al. (2020), supra). According to the present disclosure, the nucleic acid amplification reaction may be conducted using at least one of the following methodologies: polymerase chain reaction (PCR), quantitative polymerase chain reaction (qPCR), reverse transcription polymerase chain reaction (RT-PCR), reverse transcription quantitative polymerase chain reaction (RT-qPCR), nested PCR, hot-start PCR, multiplex PCR, in situ PCR, single cell PCR, touchdown PCR, ligase chain reaction (LCR), gap ligase chain reaction (gLCR) and isothermal amplification. In certain embodiments, the nucleic acid amplification reaction is an isothermal amplification reaction using any of the various natural or engineered enzymes available for this purpose, e.g., DNA polymerases having strong strand- displacement activity in isothermal conditions, thereby obviating the need for thermal cycling. Such polymerases are well-known in the art, and may include DNA polymerase long fragment (LF) of thermophilic bacteria, such as Bacillus stearothermophilus (Bst), Bacillus Smithii (Bsm), Geobacillus sp. M (GspM) and Thermodesulfatator indicus (Tin), and engineered variants therefrom as well as Taq DNA polymerase variants. Non-limiting examples of the polymerase which can be used to perform the method of the disclosure include Bst DNA polymerase, Bst 2.0 DNA polymerase, Bst 2.0 WarmStart™ DNA polymerase (New England Biolabs, Ipswich, Mass.), GspM LF DNA polymerase, GspSSD LF DNA polymerase, Tin exo-LF DNA polymerase and SD DNA polymerase Phi29 DNA polymerase, Bsu DNA polymerase, OmniAmp™ DNA polymerase (Lucigen, Middleton, Mich.), Taq DNA polymerase, Vent ^® and Deep Vent ^® DNA polymerases (New England Biolabs), 9°Nm™ DNA polymerase (New England Biolabs), Klenow fragment of DNA polymerase I, PhiPRD1 DNA polymerase, phage M2 DNA polymerase, T4 DNA polymerase, and T5 DNA polymerase, which are usually used at 50-75°C, more generally at 55 to 65°C. In some examples, about 1 to 20 U (such as about 1 to 15 U, about 2 to 12 U, about 10 to 20 U, about 2 to 10 U, or about 5 to 10 U) of a DNA polymerase is included in the reaction. In some examples, the polymerase used has strand displacement activity and lacks 5′-3′ exonuclease activity. In one non-limiting example, the DNA polymerase used is Bst 2.0 WarmStart™ DNA polymerase, for example, about 8 U of Bst 2.0 WarmStart™ DNA polymerase per reaction. Examples of the isothermal amplification may include, but are not limited to, strand-displacement amplification (SDA), rolling-circle amplification (RCA), cross-priming amplification (CPA), nucleic acid sequence-based amplification (NASBA), recombinase polymerase amplification (RPA), helicase-dependent amplification (HDA), transcription-mediated amplification, loop-mediated amplification (LAMP), and competitive annealing mediated isothermal amplification (CAMP). When the nucleic acid to be amplified or detected is RNA, a reverse transcription is conducted prior to the isothermal amplification of the target sequence. The reverse transcription may be performed using any suitable reverse transcriptase (RT). RT is well-known in the art and may include, but is not limited to, Avian Myeloblastosis Virus (AMV) RT and Moloney Murine Leukemia virus (MMLV) RT. The Reverse transcription and DNA amplification may be performed in the same reaction mixture that includes the DNA polymerase and RT enzymes applied. In addition, the method of the disclosure may use a DNA polymerase having both strong displacement activity and RT activity, such as Pyrophage 3173 DNA polymerase. In certain embodiments, the nucleic acid amplification reaction is conducted by RT-LAMP or real-time quantitative RT-LAMP (real-time qRT-LAMP), where denaturation of the DNA template is not required and thus the LAMP reaction can be conducted under isothermal conditions (e.g., ranging from 60°C to 67°C). The principle of the LAMP method is disclosed in T. Notomi et al. (2000), Nucleic acids research, 28:E63. In brief, the reaction is initiated by annealing and extension of a pair of “loop-forming” primers (i.e., forward inner primer (FIP) and backward inner primer (BIP)), followed by annealing and extension of a pair of flanking primers (i.e., forward outer primer (F3) and backward outer primer (B3)). Extension of these primers results in strand- displacement of the loop-forming elements, which fold up to form terminal hairpin-loop structures. Once these key structures have appeared, the amplification process becomes self-sustaining, and proceeds at a constant temperature in a continuous and exponential manner (rather than a cyclic manner, like PCR). Optionally, an additional pair of loop primers (i.e., forward loop primer (LF) and backward loop primer (LB)) may be included to accelerate the reaction. In certain embodiments, the first primer set for LAMP assay includes the first forward and backward outer primers, and a first forward inner primer having a nucleotide sequence of SEQ ID NO: 3 and a first backward inner primer having a nucleotide sequence of SEQ ID NO: 4. The first primer set may further include one of a first forward loop primer having a nucleotide sequence of SEQ ID NO: 5, a first backward loop primer having a nucleotide sequence of SEQ ID NO: 6, and a combination thereof to accelerate the reaction of LAMP. In certain embodiments, the second primer set for LAMP assay includes the second forward and backward outer primers, and a second forward inner primer having a nucleotide sequence of SEQ ID NO: 9 and a second backward inner primer having a nucleotide sequence of SEQ ID NO: 10. The second primer set may further include one of a second forward loop primer having a nucleotide sequence of SEQ ID NO: 11, a second backward loop primer having a nucleotide sequence of SEQ ID NO: 12, and a combination thereof to accelerate the reaction of LAMP. In certain embodiments, the third primer set for LAMP assay includes the third forward and backward outer primers, and a third forward inner primer having a nucleotide sequence of SEQ ID NO: 15 and a third backward inner primer having a nucleotide sequence of SEQ ID NO: 16. The third primer set may further include one of a third forward loop primer having a nucleotide sequence of SEQ ID NO: 17, a third backward loop primer having a nucleotide sequence of SEQ ID NO: 18, and a combination thereof to accelerate the reaction of LAMP. In certain embodiments, the fourth primer set for LAMP assay includes the fourth forward and backward outer primers, and a fourth forward inner primer having a nucleotide sequence of SEQ ID NO: 21 and a fourth backward inner primer having a nucleotide sequence of SEQ ID NO: 22. The fourth primer set may further include one of a fourth forward loop primer having a nucleotide sequence of SEQ ID NO: 23, a fourth backward loop primer having a nucleotide sequence of SEQ ID NO: 24, and a combination thereof. In certain embodiments, the fifth primer set for LAMP assay includes the fifth forward and backward outer primers, and a fifth forward inner primer having a nucleotide sequence of SEQ ID NO: 27 and a fifth backward inner primer having a nucleotide sequence of SEQ ID NO: 28. The fifth primer set may further include one of a fifth forward loop primer having a nucleotide sequence of SEQ ID NO: 29, a fifth backward loop primer having a nucleotide sequence of SEQ ID NO: 30, and a combination thereof. According to the present disclosure, the at least one primer set may further include a sixth primer set for amplifying a region of S gene of SARS- CoV-2. The sixth primer set includes a sixth forward outer primer having a nucleotide sequence of SEQ ID NO: 31 and a sixth backward outer primer having a nucleotide sequence of SEQ ID NO: 32. In certain embodiments, the sixth primer set for LAMP assay includes the sixth forward and backward outer primers, and a sixth forward inner primer having a nucleotide sequence of SEQ ID NO: 33 and a sixth backward inner primer having a nucleotide sequence of SEQ ID NO: 34. The sixth primer set may further include one of a sixth forward loop primer having a nucleotide sequence of SEQ ID NO: 35, a sixth backward loop primer having a nucleotide sequence of SEQ ID NO: 36, and a combination thereof. In certain embodiments, the reaction mixture for the LAMP reaction may further include a suitable buffer (such as a phosphate buffer or Tris buffer). The buffer may also include additional components, such as potassium and/or sodium salts (such as KCl or NaCl), magnesium and/or manganese salts (e.g., MgCl ₂ , MgSO ₄ , MnC ₁₂ , and/or MnSO ₄ ), and/or ammonium salts (e.g., (NH ₄ ) ₂ SO ₄ )), detergents (e.g., TWEEN ^® -20, TRITON ^® -X100), or other additives (such as betaine or dimethylsulfoxide). One of skill in the art can select an appropriate buffer and any additives using routine skill. In one non-limiting example, the buffer (pH 8.6) includes 50 mM KCl, 10 mM (NH4)2SO4, 0.1% TWEEN ^® -20, 4 mM MgSO4, and 0.8 M betaine. Exemplary commercially available reaction buffers include 1× Isothermal Amplification Buffer (New England Biolabs, Ipswich, Mass.), LoopAmp Reaction Mix (Eiken Chemical Co., Ltd., Tokyo, Japan), and ILLUMIGENE reaction buffer (Meridian Bioscience, Inc., Cincinnati, Ohio). The reaction mixture may also include nucleotides or nucleotide analogs. In some examples, an equimolar mixture of dATP, dCTP, dGTP, and dTTP (referred to as dNTPs) may be included. In certain embodiments, a detectable label may be attached or conjugated to the primer(s) according to this disclosure using techniques well known to those skilled in the art, so as to quantitatively detect the target SARS-CoV-2. Examples of the detectable label suitable for use in the present disclosure include, but are not limited to: a hapten label, such as biotin and digoxigenin; a chemiluminescent label, such as acridinium esters, thioesters, orsulfonamides, luminol, isoluminol, phenanthridinium esters, and the like; a fluorescent label, such as fluorescein, rhodamine, FAM, TET, HEX, JOE, TAMA, NTB, TAMRA, ROX, VIC, NED, cyanin dye, Texas Red, DABCYL, DABSYL, malachite green, AlexaFluor dye, LC-Red dye, PromoFluor dye, and derivatives thereof; a radioactive label, such as ³ H, ¹²⁵ I, ³⁵ S, ¹⁴ C, or ³² P; an enzymatic label, such as horse radish peroxidase, alkaline phosphatase, and the like; a particle label, such as gold colloids, quantum dot, etc.; and a colorimetric label, such as colored glass or plastic (e.g., polystyrene, polypropylene, latex, etc.) beads. According to this disclosure, the primers may be modified to have 15 to 35 nucleotides in length by adding at least one nucleotide residue at the 5’ and/or 3’ terminal end thereof or by deleting at least one nucleotide residue from the 5’ and/or 3’ terminal end thereof. According to this disclosure, the amplification product may be detected using any of the various methods available for this purpose which are well- known in the art. For example, the detection method may be turbidity measurement, fluorescence detection, bioluminescence detection, gel electrophoresis, colorimetric detection, immunoenzymatic detection, electrochemical detection, and combinations thereof. The detection may be semi-quantitative or quantitative. The detection may also be real-time detection, wherein the signal resulting from the presence of the amplification product is measured during the course of the nucleic acid amplification reaction to monitor the accumulation of specific amplification products. Turbidity measurement is used when the amplification reaction, such as LAMP, produces large amounts of magnesium pyrophosphate (a white precipitate) and dsDNA, which allow visual inspection of results or with a turbidimeter. Fluorescence detection may use DNA intercalating dyes, fluorescent molecular beacon probes or a fluorescence metal indicator (such as calcein). Bioluminescence detection may be conducted through measurement of bioluminescent output of the coupled conversion of inorganic pyrophosphate produced stoichiometrically during nucleic acid synthesis to ATP by the enzyme ATP sulfurylase. Colorimetric detection may use a colored indicator for alkaline metal ions, such as hydroxy naphthol blue (HNB) (see Goto et al., BioTechniques, 2009 Mar, 46(3):167-72), and/or a pH indicator with a capacity of prominent color alteration within the pH range of 7.5 ± 1.1, such as phenol red (pH 6.8- 8.2), neutral red (pH 6.8-8.0), cresol red (pH 7.2- 8.8), etc. Electrochemical detection may use a pH meter for direct measurement of released hydrogen ions during the amplification reaction, such as LAMP, or integrated electrodes for measuring decreases in current resulting from increasing binding of electrochemically-active DNA-binding redox reporters, such as Methylene Blue, to amplification products. Immunoenzymatic detection includes enzyme- linked immunosorbent assays (ELISA) and lateral flow immunoassays based on the use of specific probes (see, e.g. Hashimoto M. et al., Malar J., 2018 Jun 19, 17(1):235; and Sun Yu-Ling et al., J. Vet Med Sci., 2014 Apr, 76(4):509–516). As used herein, the “biological sample” is a sample obtained from an organism (such as an animal subject) or from components (such as cells and tissues) of the organism. Examples of the biological sample include, but are not limited to, a blood sample, a plasma sample, a serum sample, a corneal tissue sample, a tear sample, a saliva sample, a cerebrospinal fluid sample, a feces sample, a tissue biopsy, a surgical specimen, a urine sample, a fine needle aspirate, and combinations thereof. According to the present disclosure, a kit for detecting SARS-CoV-2 is provided, and includes at least one of the primer sets as described above. In certain embodiments, the kit further includes the colored indicator for alkaline metal ions, the pH indicator or the combination thereof. The kit may also further include a nucleic acid dye for monitoring the DNA amplification product. Examples of the nucleic acid dye include, but are not limited to, ethidium bromide (EtBr), SYBR GREEN I, SYBR GREEN II, SYBR Orange, SYBR GOLD, propidium iodide (PI), SYTOX Blue and SYPRO Ruby, etc. The disclosure will be further described by way of the following examples. However, it should be understood that the following examples are solely intended for the purpose of illustration and should not be construed as limiting the disclosure in practice. EXAMPLES General Experimental Materials: 1. The primers used in the following experiments were synthesized by Genomics Co., Ltd. 2. 1 kb DNA RTU Ladder (100 bp-3 k)(Cat. No. BR332- 500, Biomate ^TM ) used in the following experiments was purchased from Rainbow Biotechnology Co., Ltd. 3. The following plasmids were custom-made from GENEWIZ Inc.: (1) Plasmid p3.1-SARS2-nsp2 (1,914 bps), which carries nonstructural protein 2 (nsp2) gene of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2); (2) Plasmid p3.1-SARS1-nsp2 (1,914 bps), which carries nsp2 gene of severe acute respiratory syndrome coronavirus 1 (SARS-CoV-1); (3) Plasmid p3.1-MERS-nsp2 (1,980 bps), which carries nsp2 gene of middle east respiratory syndrome coronavirus (MERS-CoV); (4) Plasmid p3.1-OC43-nsp2 (1,815 bps), which carries nsp2 gene of human coronavirus OC43 (HCoV-OC43); (5) Plasmid p3.1-SARS2-nsp4 (1,500 bps), which carries nonstructural protein 4 (nsp4) gene of SARS-CoV-2; (6) Plasmid p3.1-SARS1-nsp4 (1,500 bps), which carries nsp4 gene of SARS-CoV-1; (7) Plasmid p3.1-MERS-nsp4 (1,521 bps), which carries nsp4 gene of MERS-CoV; (8) Plasmid p3.1-OC43-nsp4 (1,488 bps), which carries nsp4 gene of HCoV-OC43; and (9) Plasmid pcDNA3.1 (5,428 bps), which carries a cytomegalovirus (CMV) promoter. 4. The following plasmids were custom-made from GenScript: (1) Plasmid pBS-SARS2-spike (3,822 bps), which carries spike (S) gene of SARS-CoV-2; (2) Plasmid pBS-SARS1-spike (3,768 bps), which carries S gene of SARS-CoV-1; (3) Plasmid pBS-MERS-spike (4,062 bps), which carries S gene of MERS-CoV; and (4) Plasmid pBS-OC43-spike (4,062 bps), which carries S gene of HCoV-OC43. 5. The total RNA of a respective one of the six respiratory viruses listed in Table 1 was obatained from the virology laboratory of Department of Pathology, National Cheng Kung University Hospital (Tainan, Taiwan^^&KLQD).

Table 1 6. Respiratory Evaluation Panel 01 (Qnostics, Cat. No.RESPEP01-C) used in the following experiments was purchased from YUAN IN Group CO., Ltd., which includes the six respiratory viruses shown in Table 2.

Table 2 7. Source and cultivation of human lung adenocarcinoma cell line A549: Human lung adenocarcinoma cell line A549 was purchased from American Type Culture Collection (ATCC ^® CCL-185 ^TM , ATCC, Manassas, Va., USA). The A549 cells were grown in a 100-mm Petri dish containing Dulbecco’s Modified Eagle’s Medium (DMEM)(Corning) supplemented with 10% fetal bovine serum (FBS), 50 U/mL penicillin, and 100 ^g/mL streptomycin. The A549 cells were cultivated in an incubator with culture conditions set at 37°C and 5% CO2. Medium change was performed every four to five days. Cell passage was performed when the cultured cells reached 80% of confluence. General Procedures: 1. Preparation of RNA transcript mixture A respective one of the plasmid p3.1-SARS2-nsp2 and plasmid p3.1-SARS2-nsp4 described in section 3 of “General Experimental Materials” was used as a template and was subjected to in vitro transcription using RiboMAX™ Large Scale RNA Production System-T7 (Promega), so as to obtain a respective one of a RNA transcript having nsp2 gene of SARS-CoV-2 and a RNA transcript having nsp4 gene of SARS-CoV-2. Thereafter, the RNA transcript having nsp2 gene of SARS-CoV-2 and the RNA transcript having nsp4 gene of SARS-CoV-2 were mixed to obtain a RNA transcript mixture. Example 1. Primer sets for detecting SARS-CoV-2 The primer pairs and loop primer pairs for detecting SARS-CoV-2 were designed based on the conserved regions of nsp2 gene, nsp4 gene, and S gene in the complete genome sequence of SARS-CoV-2 (GenBank accession numbers NC_045512.2 and MN908947.3) using PrimerExplorer V5 software. The specificity of the primer pairs and the loop primer pairs was analyzed by BLAST analysis available in the GenBank of the NCBI website (http://www.ncbi.nlm.nih.gov/BLAST). Therefore, 5 primer sets, including primer sets nsp2-1, nsp2-2, nsp4-1, nsp4-2, and S-1, were obtained for specific detection of SARS-CoV-2. A respective one of the five primer sets included one outer primer pair, one inner primer pair, and one loop primer pair. In addition, a loop primer pair (designated as S- 2-LF / S-2-LB) was also obtained for specific detection of SARS-CoV-2, and was used in combination with an outer primer pair (designated as S-2-F3 / S- 2-B3) and an inner primer pair (designated as S-2- FIP / S-2-BIP) that were described in W.E. Huang et al. (2020), Microbial Biotechnology, 13(4):950-961. The combination of the three primer pairs is referred to as “primer set S-2” hereinafter. The detailed information of the abovementioned primer sets are summarized in Tables 3 to 8.

g c s e d i t o e l c u n d e n i l r e d n u e h t o t g n i d n o p s e r r o c s e u d i s e r e d i t o e l c u n e h t b c . s e d i t o e l c u n d e n i l r e d n u e h t o t g n i d n o p s e r r o c s e u d i s e r e d i t o e l c u n e h t : b

e d i t o e l c u n d e n i l r e d n u e h t o t g n i d n o p e r r o c e u d i e r e d i t o e l c u n e h t b

e d i t o e l c u n d e n i l r e d n u e h t o t g n i d n o p e r r o c e u d i e r e d i t o e l c u n e h t b

s e d i t o e l c u n d e n i l r e d n u e h t o t g n i d n o p s e r r o c s e u d i s e r e d i t o e l c u n e h t b c s e d i t o e l c u n d e n i l r e d n u e h t o t g n i d n o p s e r r o c s e u d i s e r e d i t o e l c u n e h t b Example 2. Evaluation for detection effect of aforesaid six primer sets on SARS-CoV-2 through loop-mediated isothermal amplification (LAMP) assay A. Specificity test A respective one of the twelve plasmids described in sections 3 and 4 of “General Experimental Materials” was used as a template and was subjected to LAMP assay that was performed using the corresponding primer set shown in Table 9 and the reaction conditions shown in Table 10. Table 9 Table 10 The color change of the reaction mix in the reaction tube was visually observed, and the reaction mix changed its color from red to yellow-orange in response to the pH change during nucleic acid amplification. The resultant product having a yellow- orange color indicated that the amplification of DNA of SARS-CoV-2 was successful. To confirm that the color change was due to target DNA amplification, the resultant products were subjected to 1.5% agarose gel electrophoresis to verify the presence of amplicons. FIGS. 1 to 6 respectively show the detection effect of the six primer sets of the present disclosure on SARS-CoV-2. As shown in FIG. 1, the gel electrophoresis analysis demonstrated that regarding the use of the primer set nsp2-1 for performing the LAMP assay, a ladder-like banding pattern was observed on the corresponding target plasmid (namely the plasmid p3.1-SARS2-nsp2). In addition, a LAMP amplified product having a yellow- orange color was visually observed on the plasmid p3.1-SARS2-nsp2. No ladder-like banding patterns and color change were observed on the non-target plasmids (namely the plasmid p3.1-SARS1-nsp2, the plasmid p3.1-MERS-nsp2, and the plasmid p3.1-OC43-nsp2). Besides, similar results were observed with respect to the primer sets nsp2-2, nsp4-1, nsp4-2, S-1, and S-2 (see FIGS. 2 to 6). These results indicate that the use of the six primer sets of the present disclosure for detecting SARS-CoV-2 through LAMP assay can exhibit high specificity. B. Sensitivity test A respective one of the plasmid p3.1-SARS2-nsp2, plasmid p3.1-SARS2-nsp4, and plasmid pBS-SARS2-spike described in sections 3 and 4 of “General Experimental Materials” was subjected to 10-fold serial dilution with DEPC-treated water, so as to obtain six DNA dilutions having different concentrations (5×10 ⁵ copies/rxn, 5×10 ⁴ copies/rxn, 5×10 ³ copies/rxn, 5×10 ² copies/rxn, 5×10 ¹ copies/rxn, and 5×10 ⁰ copies/rxn). A respective one of the six DNA dilutions of each plasmid was used as a template and was subjected to LAMP assay that was performed using the corresponding primer set shown in Table 9 and the reaction conditions shown in Table 10. The resultant products were subjected to 1.5% agarose gel electrophoresis to verify the presence of amplicons. Based on the result of the LAMP assay (data not shown), it was found that the detection limit of the respective one of the six primer sets for SARS-CoV- 2 was 5×10 ⁰ copies/rxn, indicating that the use of the six primer sets of the present disclosure for detecting SARS-CoV-2 through LAMP assay can exhibit high sensitivity. Example 3. Evaluation for detection effect of aforesaid six primer sets on SARS-CoV-2 through reverse transcription loop- mediated isothermal amplification (RT- LAMP) assay The total RNA of SARS-CoV-2 (German strain) described in section 5 of “General Experimental Materials” was subjected to 10-fold serial dilution with DEPC-treated water, so as to obtain six RNA dilutions having different concentrations (5×10 ⁵ copies/rxn, 5×10 ⁴ copies/rxn, 5×10 ³ copies/rxn, 5×10 ² copies/rxn, 5×10 ¹ copies/rxn, and 5×10 ⁰ copies/rxn). A respective one of the six RNA dilutions was used as a template and was subjected to RT-LAMP assay that was performed using the six primer sets shown in Tables 3 to 8 and the reaction conditions shown in Table 11. Table 11

The resultant products were subjected to 1.5% agarose gel electrophoresis to verify the presence of amplicons. The result is shown in Table 12 below. It can be seen from Table 12 that the detection limit of the primer set S-1 for SARS-CoV-2 was 5×10 ² copies/rxn, the detection limit of the respective one of the primer sets nsp2-1, nsp4-1, and S-2 for SARS-CoV-2 was 5×10 ¹ copies/rxn, and the detection limit of the respective one of the primer sets nsp2-2 and nsp4-2 for SARS-CoV-2 was 5×10 ⁰ copies/rxn. This result suggests that the use of the six primer sets of the present disclosure for detecting SARS-CoV-2 through RT-LAMP assay can exhibit high sensitivity (especially the primer sets nsp2-2 and nsp4-2). Table 12 Example 4. Evaluation for detection effect of combined use of aforesaid primer sets on SARS-CoV-2 A. Specificity test A549 cells were divided into 6 groups, including a negative control group, a blank control group, and four experimental groups (i.e., experimental groups 1 to 4). The respective group of the A549 cells was incubated in a 35-mm Petri dish containing 3 mL of DMEM (Corning)(supplemented with 10% fetal bovine serum (FBS)) at 2.7×10 ⁵ cells/dish, followed by cultivation in an incubator (37°C, 5% CO ₂ ) for 24 hours. Afterwards, the respective one of the cell cultures of the four experimental groups was co- transfected with 2 μg of a combination of two specific plasmids as shown in Table 13 using 2 μL of TurboFect transfection reagent (Cat. No. R5031, Thermo Fisher Scientific Inc.). In addition, the cell culture of the negative control group was transfected with 2 μg of the plasmid pcDNA3.1 described in section 3 of “General Experimental Materials” using 2 μL of TurboFect transfection reagent (Cat. No. R5031, Thermo Fisher Scientific Inc.), and the cell culture of the blank control group received no treatment. Thereafter, the cell culture of the respective group was cultivated in an incubator (37°C, 5% CO2) for 6 hours. After medium change with 3 mL of a fresh DMEM (supplemented with 10% FBS, 50 U/mL penicillin, and 100 ^g/mL streptomycin), the cell culture of the respective group was cultivated in an incubator (37°C, 5% CO2) for 18 hours.

Table 13 Note: The copy number ratio of the two specific plasmids used for the respective experimental group is 1:1. The resultant culture of the respective group was subjected to extraction of RNA using Direct-zol ^TM RNA MiniPrep (Cat. No. R2052, Zymo Research) in accordance with the manufacturer’s instructions. The resultant total RNA of the respective group was used as a template and was subjected to RT-LAMP assay that was performed using the primer sets nsp2-2 and nsp4- 2 and the reaction conditions shown in Table 14. Table 14 The resultant products were subjected to colorimetric assay and agarose gel electrophoresis analysis according to the method described in Example 2. As shown in FIG. 7, the gel electrophoresis analysis demonstrated that regarding the use of a combination of the primer sets nsp2-2 and nsp4-2 for performing the RT-LAMP assay, a ladder-like banding pattern was observed on the experimental group 1. In addition, an RT-LAMP amplified product having a yellow-orange color was visually observed on the experimental group 1. No ladder-like banding patterns and color change were observed on the experimental groups 2 to 4. These results indicate that the primer sets of the present disclosure, when used in combination to detect SARS-CoV-2, can exhibit high specificity. B. Sensitivity test Three plasmid mixtures (i.e., plasmid mixtures 1 to 3) were prepared using the recipe shown in Table 15.

Table 15 Note: The copy number ratio of the two plasmids used for the respective plasmid mixture is 1:1. A respective one of the three plasmid mixtures was subjected to 10-fold serial dilution with DEPC- treated water, so as to obtain six DNA dilutions having different concentrations (5×10 ⁵ copies/rxn, 5×10 ⁴ copies/rxn, 5×10 ³ copies/rxn, 5×10 ² copies/rxn, 5×10 ¹ copies/rxn, and 5×10 ⁰ copies/rxn). A respective one of the six DNA dilutions of each plasmid mixture was used as a template and was subjected to LAMP assay that was performed using the corresponding combination of primer sets shown in Table 16 and the reaction conditions shown in Table 17. Table 16 Table 17 The resultant products were subjected to 1.5% agarose gel electrophoresis to verify the presence of amplicons. Based on the result of the LAMP assay (data not shown), it was found that the detection limit of the respective one of the three combinations of primer sets (i.e., S-1/nsp4-1, S-1/nsp4-2, and nsp2-2/nsp4- 2) for SARS-CoV-2 was 5×10 ⁰ copies/rxn, indicating that the primer sets of the present disclosure, when used in combination to detect SARS-CoV-2, can exhibit high sensitivity. Example 5. Evaluation for detection effect of combined use of primer sets nsp2-2 and nsp4-2 on SARS-CoV-2 and other respiratory viruses Experimental procedures: The RNA transcript mixture described in section 1 of “General Procedures” and the total RNA of a respective one of IAV, IBV, HPIV, RSV, and ADV described in section 5 of “General Experimental Materials” were respectively used as a template and were respectively subjected to RT-LAMP assay that was performed using the primer sets nsp2-2 and nsp4- 2 and the reaction conditions shown in Table 14. In addition, a respective one of the six respiratory viruses described in section 6 of “General Experimental Materials” was subjected to extraction of RNA using QIAamp Viral RNA Mini kit (QIAGEN) in accordance with the manufacturer’s instructions. The resultant total RNA of each virus and the RNA transcript mixture described in section 1 of “General Procedures” were respectively used as a template and were respectively subjected to RT- LAMP assay that was performed using the primer sets nsp2-2 and nsp4-2 and the reaction conditions shown in Table 14. The total RNA extracted from A549 cells that were transfected with the plasmid pcDNA3.1 described in section 3 of “General Experimental Materials” was used as negative control. The resultant products were subjected to colorimetric assay and a 1.5% agarose gel electrophoresis analysis according to the method described in Example 2. Results: As shown in FIGS. 8 and 9, the gel electrophoresis analysis demonstrated that regarding the use of the combination of the primer sets nsp2-2 and nsp4-2 for performing the RT-LAMP assay, a ladder-like banding pattern was observed on SARS-CoV-2. In addition, an RT-LAMP amplified product having a yellow-orange color was visually observed on SARS-CoV-2. No ladder- like banding patterns and color change were observed on the other respiratory viruses. These results indicate that the primer sets nsp2- 2 and nsp4-2 of the present disclosure, when used in combination to detect SARS-CoV-2, can exhibit high specificity. Example 6. Evaluation for detection effect of combined use of primer sets nsp2-2 and nsp4-2 on different SARS-CoV-2 strains Experimental procedures: The total RNA of a respective one of the three SARS-CoV-2 strains described in section 5 of “General Experimental Materials” was subjected to 10-fold serial dilution with DEPC-treated water, so as to obtain seven RNA dilutions having different concentrations (5×10 ⁶ copies/rxn, 5×10 ⁵ copies/rxn, 5×10 ⁴ copies/rxn, 5×10 ³ copies/rxn, 5×10 ² copies/rxn, 5×10 ¹ copies/rxn, and 5×10 ⁰ copies/rxn). A respective one of the seven RNA dilutions was used as a template and was subjected to real-time quantitative RT-LAMP (real-time qRT-LAMP) assay that was performed on a StepOnePlus™ real-time PCR system (Applied Biosystems) using the primer sets nsp2-2 and nsp4-2 and the reaction conditions shown in Table 18.

Table 18 Results: The result is shown in Table 19 below. It can be seen from Table 19 that the detection limit of the combination of the primer sets nsp2-2 and nsp4-2 for each SARS-CoV-2 strain was 5×10 ¹ copies/rxn, and the averaged maximum cycle threshold (Ct) values of the SARS-CoV-2 German strain, UK strain, and French strain were 41.27±4.42, 43.48±2.57, and 40.49±3.10 (mean ± SD), respectively. As each amplification cycle was performed for 1 minute, indicating that a respective one of the three SARS-CoV-2 strains at a concentration of 50 RNA copies/rxn can be detected within 40 to 45 minutes. Based on the aforesaid results, the applicant suggests that the maximum cut- off Ct value that equals to 55 minutes (which is determined as the upper limit of the maximum confidence interval (99.7%) of the normal distribution within the mean plus and minus three standard deviations) can be used to determine the positivity or negativity of a given RNA specimen. When the Ct value is less than or equal to 55 minutes, the test result is assessed as positive. When the Ct value is greater than 55 minutes, the test result is assessed as negative. Summarizing the above test results, the primer sets nsp2-2 and nsp4-2 of the present disclosure, when used in combination to detect SARS-CoV-2, can exhibit high sensitivity, and are suitable for clinically predicting viral load. Table 19 While the disclosure has been described in connection with what are considered the exemplary embodiments, it is understood that this disclosure is not limited to the disclosed embodiments but is intended to cover various arrangements included within the spirit and scope of the broadest interpretation so as to encompass all such modifications and equivalent arrangements.

SEQUENCE LISTING

<110> NATIONAL CHENG KUNG UNIVERSITY

TAIWAN CARBON NANO TECHNOLOGY CORPORATION

<120> METHODS AND KITS FOR DETECTING SEVERE ACUTE RESPIRATORY

SYNDROME CORONAVIRUS 2

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