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. 2023 Nov;102(11):103027.
doi: 10.1016/j.psj.2023.103027. Epub 2023 Aug 14.

Rapid detection of pigeon Megrivirus using TaqMan real-time PCR technology

Jinpeng Zhang  1 , Huanru Fu  2 , Cuiteng Chen  1 , Jinxiu Jiang  1 , Yusheng Lin  1 , Bin Jiang  1 , Lin Lin  1 , Qilin Hu  1 , Chunhe Wan  3
Affiliations

Rapid detection of pigeon Megrivirus using TaqMan real-time PCR technology

Jinpeng Zhang et al. Poult Sci. 2023 Nov.

Abstract

Megriviruses have been identified from fecal samples in wild pigeons in Hong Kong (China) and Hungary. In this study, the genomic sequences of pigeon Megriviruses (PiMeVs) were downloaded from GenBank and compared. Based on the genetic comparison results, a pair of primers and TaqMan probe were designed based on the conserved sequences of the 3C gene (located in the P3 gene coding region), and a TaqMan real-time PCR method (TaqMan-qPCR) was established. The standard curve of the TaqMan-qPCR had an axial intercept of 39.74 and a slope of -3.2475 with a linear correlation (R2) of 1.00 and an efficiency of 103.2%. No cross-amplification signal was found from other pigeon viruses (such as avian influenza virus, pigeon paramyxovirus type I, pigeon torque teno virus, pigeon adenovirus, and pigeon circovirus). The limit of detection concentration was 53.6 copies/μL. The intra- and interassay results were less than 1.0% based on the reproducibility test. Furthermore, field samples investigation by the established TaqMan-qPCR method showed that positive signals can be found from racing pigeon fecal samples and embryos. Thus, our data suggested that this visible TaqMan-qPCR method is sensitive, specific, and reproducible. Moreover, we first confirmed the presence of pigeon Megrivirus infection in racing pigeon embryos, indicating that the virus may be vertically transmitted. This study provides a reference basis for further understanding the epidemiology of PiMeVs.

Keywords: Megrivirus; TaqMan-qPCR; racing pigeons; vertical transmission.

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Figures

Figure 1
Figure 1
Standard curve of the TaqMan-qPCR method.
Figure 2
Figure 2
Specificity test of the TaqMan-qPCR method 1: PiMeV; Controls: AIV, PPMV-1, PTVV, PiAd, PiCV, and ddH2O. These controls were all found to have no positive signal. It could not be effectively distinguished by the naked eye.
Figure 3
Figure 3
Sensitivity test of the TaqMan-qPCR method 1-3: serial 10-fold dilutions of plasmid DNA (5.36 × 102–5.36 × 100 copies/μL); 4: negative control.
See this image and copyright information in PMC

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