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Updated: May 15, 2026
Detecting SARS-CoV-2 Virus by Reverse Transcription-Loop-Mediated Isothermal Amplification
Published on: September 8, 2023
Development of a loop-mediated isothermal amplification method to rapidly detect porcine circovirus genotypes 2a and
Xiaohuo Qiu1, Tian Li, Guorui Zhang
1Key Laboratory of Animal Virology of Ministry of Agriculture, College of Animal Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, PR China.
Background:
Porcine circovirus type 2 (PCV2), is nowadays associated with a number of diseases known as porcine circovirus-associated diseases (PCVAD), especially postweaning multisystemic wasting syndrome (PMWS). The epidemiological investigation of PCV2 infection was usually conducted by PCR, nested PCR, PCR-RFLP, TaqMan-based assay and nucleotide sequencing. However, there is still no rapid, sensitive and practical method for detecting PCV2 genotypes. As a novel nucleic acid amplification method, the loop-mediated isothermal amplification method (LAMP) has been used to detect a variety of pathogenic microorganisms.
Results:
Herein, a LAMP method is developed to detect the genotypes of PCV2. The diagnostic sensitivity of LAMP is 1 copy/reaction for differentiating genotypes PCV2a and PCV2b. The reaction process was completed at 65°C for 1 hour in a water bath. Cross-reactivity assay shows that this method is specific for PCV2a and PCV2b and no reactive for PCV2c and other swine-origin viruses (i.e. CSFV, PRRSV, BVDV, TGEV and PEDV, etc). Identity between LAMP and nested PCR was 92.3% on 52 field clinical samples.
Conclusions:
LAMP method provides a rapid, sensitive, reliable way to detect PCV2a and PCV2b, and a better means for the large scale investigation of PCV2a and PCV2b infection.
