Establishment of triple chip digital PCR method for human herpesvirus 6
10.3760/cma.j.cn112866-20210718-00129
- VernacularTitle:人疱疹病毒6型三重芯片式数字PCR方法的建立
- Author:
Wenjun WANG
1
;
Juan SONG
;
Ruifang WANG
;
Yiqiu WAN
;
Ze WEI
;
Hailan YAO
;
Jun HAN
Author Information
1. 中国疾病预防控制中心病毒病预防控制所 传染病预防控制国家重点实验室，北京 102206
- Keywords:
Human herpesvirus 6;
Triple chip digital PCR;
Chromosomal integrated HHV-6
- From:
Chinese Journal of Experimental and Clinical Virology
2021;35(5):570-574
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To determine the viral load of human herpesvirus 6 A (HHV-6A), HHV-6B and chromosomal integrated HHV-6 (ciHHV-6) simultaneously through a triple chip digital PCR (tcdPCR) method for detection of HHV-6A/6B and ribonuclease P-30 (RPP30).Methods:According to optimal reaction conditions of real-time fluorescence quantitative PCR (RT-qPCR) method, the tcdPCR mehod of HHV-6A, HHV-6B and RPP30 was established. The sensitivity of tcdPCR was determined by virus cultures and the specificity of tcdPCR was detected with other herpesviruses. Subsequently, the tcdPCR of HHV-6A, HHV-6B and RPP30 was verified through 127 whole blood samples.Results:The consistency between RT-qPCR and tcdPCR for HHV-6 detection was good (R 2>0.97). And there was no cross-reaction with other herpesviruses. The 14 positive samples could be detected effectively by the tcdPCR of HHV-6A, HHV-6B and RPP30. The lowest detectable viral load of HHV-6A and HHV-6B was 50 copies/ml and 105 copies/ml, respectively. And the ratio of HHV-6/(RPP30/2) in 14 positive samples was less than 1. Conclusions:The tcdPCR has good sensitivity and specificity. And HHV-6 tcdPCR method can quantitatively detect the viral load of HHV-6 infection and the copy number of RPP30, and ciHHV-6 can be judged by ratio of HHV-6/(RPP30/2) in clinical samples.
