Establishment and evaluation of Y-type primer quadruple fluorescence quantitative RT-PCR method for dengue virus
10.3760/cma.j.cn112866-20201113-00286
- VernacularTitle:登革病毒的Y型引物四重荧光定量RT-PCR方法的建立与评价
- Author:
Lei SHI
1
;
Yunqing XU
;
Yuan XU
Author Information
1. 深圳国际旅行卫生保健中心，深圳 518045
- Keywords:
Y-type primer;
Quadruple fluorescent quantitative RT-PCR;
Dengue virus;
Genotyping
- From:
Chinese Journal of Experimental and Clinical Virology
2022;36(1):111-115
- CountryChina
- Language:Chinese
-
Abstract:
Objective:This study aimed to establish a fourfold fluorescent quantitative RT-PCR detection method for rapid typing of Dengue virus (DENV) using Y-type primers and fluorescent quantitative RT-PCR technology.Methods:Four types of DENV were used to transcribe RNA in vitro to evaluate the sensitivity and specificity of the method and to verify it with clinical samples.Results:The minimum detection limits of DENV type I, II, III and IV were 5.01 copies/PCR, 6.16 copies/PCR, 6.35 copies/PCR, 6.39 copies/PCR, respectively and had no significant difference compared with single-plex real-time fluorescent quantitative RT-PCR. There was also no crossover with other viruses. The positive agreement rate and negative agreement rate of clinical specimens were both 100%.Conclusions:The Y-type primer quadruple fluorescence quantitative RT-PCR method established in this study could effectively avoid cross-reactions between different DENV types, and has good sensitivity and specificity, and can be used for rapid typing of DENV in relevant clinical specimens with high detection efficiency and low costs.
