Establishment of droplet digital PCR detection method for mpox virus
10.3760/cma.j.cn112866-20240226-00030
- VernacularTitle:猴痘病毒微液滴数字PCR检测方法的建立
- Author:
Junxia FENG
1
;
Jing YUAN
Author Information
1. 首都儿科研究所细菌学研究室，北京 100020
- Keywords:
Mpox virus;
Droplet digital PCR;
Real time fluorescence quantitative PCR
- From:
Chinese Journal of Experimental and Clinical Virology
2024;38(3):337-342
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a droplet digital polymerase chain reaction (ddPCR) detection method for monkeypox(mpox) virus.Methods:Specific primers and probe targeting the F3 L gene of mpox virus were used to develop a ddPCR detection method for the mpox virus′s nucleic acid. The ddPCR reaction conditions were optimized, and the specificity, sensitivity, and repeatability of ddPCR were assessed. Additional examination of clinical samples obtained from human pus swabs was performed and the ddPCR method′s ability to detect clinical samples was evaluated. Results:The result demonstrated that the mpox virus ddPCR detection method was established and the ddPCR reaction temperature and primer probe concentration has been optimized. The method had a minimal detection limit of 2 copies/μl and did not exhibit cross reaction with the remaining 8 viruses. Ten clinical samples of human skin pus swabs were tested for the mpox virus; five of the samples were tested positive by ddPCR, and four of the samples were tested positive by real time fluorescence quantitative PCR (qPCR).Conclusions:The established ddPCR for mpox virus is a fast, specific, and highly sensitive detection method, and can be used for the detection of mpox virus in clinical samples.
