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01.12.2017 | Methodology | Ausgabe 1/2017 Open Access

Virology Journal 1/2017

Development of real-time and lateral flow dipstick recombinase polymerase amplification assays for rapid detection of goatpox virus and sheeppox virus

Zeitschrift:
Virology Journal > Ausgabe 1/2017
Autoren:
Yang Yang, Xiaodong Qin, Xiangle Zhang, Zhixun Zhao, Wei Zhang, Xueliang Zhu, Guozheng Cong, Yanmin Li, Zhidong Zhang
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​s12985-017-0792-7) contains supplementary material, which is available to authorized users.

Abstract

Background

Goatpox virus (GTPV) and sheeppox virus (SPPV), which belong to the Capripoxvirus (CaPV), are economically important pathogens of small ruminants. Therefore, a sensitive, specific and rapid diagnostic assay for detection of GTPV and SPPV is necessary to accurately and promptly control these diseases.

Methods

Recombinase polymerase amplification (RPA) assays combined with a real-time fluorescent detection (real-time RPA assay) and lateral flow dipstick (RPA LFD assay) were developed targeting the CaPV G-protein-coupled chemokine receptor (GPCR) gene, respectively.

Results

The sensitivity of both CaPV real-time RPA assay and CaPV RPA LFD assay were 3 × 102 copies per reaction within 20 min at 38 °C. Both assays were highly specific for CaPV, with no cross-reactions with peste des petits ruminants virus, foot-and-mouth disease virus and Orf virus. The evaluation of the performance of these two assays with clinical sample (n = 107) showed that the CaPV real-time RPA assay and CaPV RPA LFD assay were able to specially detect SPPV or GTPV present in samples of ovine in liver, lung, kidney, spleen, skin and blood.

Conclusions

This study provided a highly time-efficient and simple alternative for rapid detection of GTPV and SPPV.
Zusatzmaterial
Additional file 1: Table S1. Comparison of CaPV real-time RPA assay and CaPV RPA LFD assay with real-time qPCR assay on spiked samples. (DOCX 18 kb)
12985_2017_792_MOESM1_ESM.docx
Additional file 2: Figure S1. The detection limit of CaPV RPA LFD assay. This assay was performed using a dilution series of the SPPV/Gulang 2009 genomic DNA, and NC represents negative control. (PPTX 393 kb)
12985_2017_792_MOESM2_ESM.pptx
Literatur
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