A study on the single -tube multiplex RT -real time PCR assay in detecting rotavirus,astrovirus and hepatitis A virus
- VernacularTitle:轮状病毒星状病毒和甲型肝炎病毒的单管多重荧光定量 RT －PCR 检测方法研究
- Author:
De-Shun XU
1
;
Xiao-Juan ZHU
;
Li-Ping CHEN
;
Xiao-Fang WU
;
Lei JI
Author Information
1. 湖州市疾病预防控制中心
- Keywords:
Astrovirus;
Rotavirus;
Hepatitis A virus;
Reverse transcriptase polymerase chain reaction
- From:
Journal of Preventive Medicine
2015;(4):358-361
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop and evaluate a single -tube multiplex RT -real time PCR assay for the simultaneous detection of rotavirus,astrovirus and hepatitis A virus.Methods Gen -bank sequences of rotavirus,astrovirus and hepatitis A virus were included as reference sequences.Primers and probes were designed based on the reference sequences.A multiplex real -time RT -PCR assay was developed,and the reproducibility,specificity and sensitivity of the assay were evaluated.Fecal samples from 1 28 patients with viral diarrhea were detected and verified by gene sequencing.Results There was high reproducibility and specificity of the single -tube multiplex real -time RT -PCR assay for detecting rotavirus,astrovirus and hepatitis A virus.Detection limits of 1 01 copies were achieved in tests of rotavirus,1 02 copies for astrovirus and hepatitis A virus in one reaction.3.1 3% and 1 7.97% of 1 28 clinical specimens were detected positive for astrovirus RNA and rotavirus RNA respectively.And the positive samples were verified by gene sequencing.Conclusion Rotavirus,astrovirus and hepatitis A virus can be detected and identified by the single -tube multiplex RT -real time PCR assay with high specificity and sensitivity.The assay developed in this study can be applied to the clinical diagnosis and epidemiological investigation.
