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Genotype-specific fluorogenic RT-PCR assays for the detection and quantitation of canine coronavirus type I and type II RNA in faecal samples of dogs.

Abstract：

:Two genotype-specific fluorogenic RT-PCR assays were developed for the detection and quantitation of canine coronavirus (CCoV) type I and type II RNA in the faeces of dogs with diarrhoea. Both the fluorogenic assays showed high specificity, sensitivity and reproducibility, allowing a precise quantitation of CCoV type I and type II RNA over a linear range of about eight orders of magnitude (from 10(1) to 10(8) copies of standard RNA). Comparison with genotype-specific gel-based RT-PCR assays revealed that the fluorogenic assays were more sensitive and more rapid than conventional amplifications, with a large increase in throughput. The genotype-specific fluorogenic assays were then used to detect and measure viral loads in the faecal samples collected from dogs naturally or experimentally infected with type I, type II, or both genotypes. Of 174 samples collected from naturally infected dogs, 77 were positive for CCoV type I and 46 for CCoV type II. Thirty-eight dogs were found to be infected naturally by both genotypes, with viral RNA titres generally higher for type I in comparison to type II. At the same time, dogs infected experimentally shed type I RNA with higher titres with respect to type II.

journal_name

J Virol Methods

journal_title

Journal of virological methods

authors

Decaro N,Martella V,Ricci D,Elia G,Desario C,Campolo M,Cavaliere N,Di Trani L,Tempesta M,Buonavoglia C

doi

10.1016/j.jviromet.2005.06.005

subject

Has Abstract

pub_date

2005-12-01 00:00:00

pages

72-8

issue

1-2

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(05)00198-9

journal_volume

130

pub_type

杂志文章

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