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.
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.
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DOI：
10.1016/j.jviromet.2005.06.005
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发表时间：
2005-12
影响因子：
3.1
通讯作者：
Buonavoglia C
中科院分区：
文献类型：
--
作者：
Decaro N;Martella V;Ricci D;Elia G;Desario C;Campolo M;Cavaliere N;Di Trani L;Tempesta M;Buonavoglia C
两种基因型特异性荧光RT-PCR检测和定量犬冠状病毒（CCoV）的I型和II型RNA的腹泻狗的粪便中。两种荧光检测均显示出高特异性、灵敏度和重现性，允许在约8个数量级的线性范围内（从101至108个拷贝的标准RNA）精确定量CCoV I型和II型RNA。与基因型特异性凝胶RT-PCR检测相比，荧光检测比常规扩增更灵敏、更快速，通量大幅提高。然后使用基因型特异性荧光检测法检测和测量从自然或实验感染I型、II型或两种基因型的犬中采集的粪便样本中的病毒载量。在从自然感染的狗收集的174个样品中，77个对CCoV I型呈阳性，46个对CCoV II型呈阳性。38只狗被发现自然感染这两种基因型，与II型相比，I型的病毒RNA滴度通常更高。与此同时，实验感染的狗脱落的I型RNA滴度高于II型。
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 101 to 108 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.
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影响因子：
3.1
作者：
Pratelli A;Martella V;Pistello M;Elia G;Decaro N;Buonavoglia D;Camero M;Tempesta M;Buonavoglia C
通讯作者：
Buonavoglia C
影响因子：
1.2
作者：
HOHDATSU, T;OKADA, S;KOYAMA, H
通讯作者：
KOYAMA, H
影响因子：
3.1
作者：
Pratelli, A;Elia, G;Buonavoglia, C
通讯作者：
Buonavoglia, C
影响因子：
3.1
作者：
Pratelli A;Buonavoglia D;Martella V;Tempesta M;Lavazza A;Buonavoglia C
通讯作者：
Buonavoglia C
影响因子：
3.1
作者：
Decaro N;Pratelli A;Campolo M;Elia G;Martella V;Tempesta M;Buonavoglia C
通讯作者：
Buonavoglia C