TaqMan MGB Probe Fluorescence Real-Time Quantitative PCR for Rapid Detection of Chinese Sacbrood Virus

PLoS One

Song Yingjin¹  Ma Mingxiao²  Li Li²  Li Yongfei²  Liu Jinhua³ 

[1] Agriculture and Biology Engineering College, Tianjin University, Tianjin, China;Department of Laboratory Animal Center, Liaoning Medical University, Jinzhou, China;Jilin Entry-Exit Inspection and Quarantine Bureau, Changchun, China

关键词: Polymerase chain reaction;    Reverse transcriptase-polymerase chain reaction;    Larvae;    Bees;    DNA;    Electron microscopy;    Honey bees;    Invertebrate genomics;   

DOI  :  10.1371/journal.pone.0052670

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

Sacbrood virus (SBV) is a picorna-like virus that affects honey bees (Apis mellifera) and results in the death of the larvae. Several procedures are available to detect Chinese SBV (CSBV) in clinical samples, but not to estimate the level of CSBV infection. The aim of this study was develop an assay for rapid detection and quantification of this virus. Primers and probes were designed that were specific for CSBV structural protein genes. A TaqMan minor groove binder (MGB) probe-based, fluorescence real-time quantitative PCR was established. The specificity, sensitivity and stability of the assay were assessed; specificity was high and there were no cross-reactivity with healthy larvae or other bee viruses. The assay was applied to detect CSBV in 37 clinical samples and its efficiency was compared with clinical diagnosis, electron microscopy observation, and conventional RT-PCR. The TaqMan MGB-based probe fluorescence real-time quantitative PCR for CSBV was more sensitive than other methods tested. This assay was a reliable, fast, and sensitive method that was used successfully to detect CSBV in clinical samples. The technology can provide a useful tool for rapid detection of CSBV. This study has established a useful protocol for CSBV testing, epidemiological investigation, and development of animal models.

【 授权许可】

CC BY