Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Apr;12(4):91.
doi: 10.1007/s13205-022-03150-1. Epub 2022 Mar 14.

Establishment of SYBR green I-based quantitative real-time polymerase chain reaction for the rapid detection of a novel Chaphamaparvovirus in cats

Xunbi Liu  1 , Shuyan Li  1 , Xuan Liu  1 , Run Wang  1 , Xiangyu Xie  1 , Haiqiang Wu  1 , Yong Wang  1
Affiliations

Establishment of SYBR green I-based quantitative real-time polymerase chain reaction for the rapid detection of a novel Chaphamaparvovirus in cats

Xunbi Liu et al. 3 Biotech. 2022 Apr.

Abstract

Feline parvovirus causes infectious diseases, and Chaphamaparvovirus is a novel type of feline parvovirus. The present study aims to establish a method that can be used in clinical rapid detection of feline Chaphamaparvovirus (FeChPV), for facilitate the timely and effective diagnosis and treatment of sick animals and shorten the diagnosis time of clinical diseases. The experimental samples in this study are from 20 cats undergoing physical examination in Hefei Xin'an Animal Hospital. An SYBR Green I-based qPCR assay was performed to detect FeChPV. A pair of specific primers was designed based on the VP1 gene to perform the assay. The detection assay showed high sensitivity with a detection limit of 1.07 × 101 copies/μL and high specificity for detection of only the target virus. The coefficients of C t value variation were calculated to assess the reproducibility of the qPCR assay, and the inter- and intra-assay ranged from 0.21 to 0.67% and 0.10 to 0.56%, respectively. The result of clinical sample detection showed that the infection rate of FeChPV in 124 samples detected using qPCR assay was higher than that with conventional PCR. The established qPCR assay could be a low-cost, convenient, and reliable method to detect FeChPV in clinical practice.

Keywords: Detection; Feline Chaphamaparvovirus; Parvovirus; SYBR Green I real-time qPCR; VP1 gene.

PubMed Disclaimer

Conflict of interest statement

Conflict of interestAll authors have declared that there was no competing interests existing in the study.

Figures

Fig. 1
Fig. 1
a Amplification curve of SYBR Green I-based quantitative real-time polymerase chain reaction (PCR). The copy numbers of feline Chaphamaparvovirus (FeChPV) DNA ranged from 1.07 × 108 to 1.07 × 101 copies/μL. b Standard curve of FeChPV (concentrations ranging from 1.07 × 108 to 1.07 × 101 copies/μL, y =  − 3.330 ×  + 34.166, R2 = 0.999, E = 99.7%). c Melting curve of FeChPV (Tm = 79 ℃). d The gel electropherogram result of conventional PCR. DNA marker of 2000 bp was used. Lanes 1–8 underwent tenfold serial dilutions of the plasmids ranging from 1.07 × 108 to 1.07 × 101 copies/μL; Lane 9 was a negative control. The lowest limit of detection of conventional PCR was 1.07 × 103 copies/μL
Fig. 2
Fig. 2
Sensitivity analysis. Amplification curve of SYBR Green I-based real-time polymerase chain reaction assay of feline Chaphamaparvovirus. The lowest limit of detection of the assay was 1.07 × 101 copies/μL
Fig. 3
Fig. 3
Specificity analysis. There is no specific curve of feline astrovirus, feline bocavirus, feline panleukopenia virus, feline calicivirus, feline herpesvirus, feline coronavirus, and double-distilled water
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Chang WS, Li CX, Hall J, Eden JS, Hyndman TH, Holmes EC, et al. Meta-transcriptomic discovery of a divergent circovirus and a chaphamaparvovirus in captive reptiles with proliferative respiratory syndrome. Viruses. 2020;12:108203. - PMC - PubMed
    1. Cotmore SF, Agbandje-McKenna M, Chiorini JA, Mukha DV, Pintel DJ, Qiu J, Soderlund-Venermo M, et al. The family Parvoviridae. Adv Virol. 2014;159:1239–1247. - PMC - PubMed
    1. Di Martino B, Lanave G, Di Profio F, Melegari I, Marsilio F, Camero M, et al. Identification of feline calicivirus in cats with enteritis. Transbound Emerg Dis. 2020;67:2579–2588. doi: 10.1111/tbed.13605. - DOI - PubMed
    1. Di Profio F, Sarchese V, Palombieri A, Fruci P, Massirio I, Martella V, et al. Feline chaphamaparvovirus in cats with enteritis and upper respiratory tract disease. Transbound Emerg Dis. 2021 doi: 10.1111/tbed.14032. - DOI - PubMed
    1. Du J, Wang W, Chan JF, Wang G, Huang Y, Yi Y, et al. Identification of a novel ichthyic parvovirus in marine species in Hainan Island, China. Front Microbiol. 2019;10:2815. doi: 10.3389/fmicb.2019.02815. - DOI - PMC - PubMed

LinkOut - more resources