What is the clinical evidence from this study?

Study design: Other. Population: African swine fever virus (ASFV). Intervention: Triplex real-time PCR method vs. Other real-time PCR methods. Primary outcome: Detection limit.

September 15, 2022Frontiers in Veterinary Science39 citationsOpen Access

A triplex real-time PCR method to detect African swine fever virus gene-deleted and wild type strains

HYHao YangYango University ZPZhong PengHuazhong Agricultural University WSWenbo SongNanjing Normal University

Key Result

The developed triplex real-time PCR method successfully detected and differentiated African swine fever virus gene-deleted and wild-type strains with detection limits of 78.9, 47.0, and 82.1 copies/μl for B646L, MGF_360-14L, and CD2v genes, respectively.

Structured PICO

Population

Synthesized standard plasmids and genomic DNA from African swine fever virus (ASFV) and other swine viruses (PRV, PRRSV, JEV, PPV, PCV2)

Intervention

Triplex real-time PCR method targeting B646L, MGF_360-14L, and CD2v genes

Comparator

Reported triplex real-time PCR method (r-PCR) and recommended real-time PCR method in China (gb-PCR)

Outcome

Detection limit, specificity, and sensitivity of the PCR methodsurrogate

A novel triplex real-time PCR method was developed with high specificity and sensitivity to detect and differentiate ASFV gene-deleted and wild type strains.

Limitations

Other genes have been demonstrated as suitable targets for deletion to develop vaccine candidates, which should be included in future multiplex real-time PCR methods.

Abstract

Currently there is still no effective vaccines and drugs available for African swine fever virus (ASFV), a life-threatening virus to domestic pigs and wild boars. Therefore, accurate diagnosis is important for the prevention and control of the virus. In this study, we developed a triplex real-time PCR method to detect and differentiate ASFV gene-deleted and wild type strains based on three viral genes B646L, MGF₃60-14L gene, and CD2v. Standard curves plotted showed that there was a strong linear correlation (R 2 0. 99) between Ct values and the corresponding copy numbers of synthesized standard plasmids. The detection limits of the method for B646L, MGF₃60-14L, and CD2v were 78. 9, 47. 0, and 82. 1 copies/μl, respectively. Detection results of different types of swine viruses showed that the method only gave amplification curves to ASFV. Finally, we found the triplex real-time PCR method developed in this study displayed better results on detecting the laboratory sample mocks, and it could be used as a supplemental method to detect ASFV genotype I strains. These findings suggest that the triplex real-time PCR method developed in this study have good specificity and sensitivity. This triplex real-time PCR method might also represent an effective tool for the detection of ASFV gene-deleted and wild type strains.

问 AI

Bookmark

View Full Paper