用套式PCR方法检出腹泻及健康犬粪中的犬冠状病毒

Canine Coronavirus Detection in Feces from Diarrhea and Healthy Dogs by Nested-PCR

自2003年夏至2004年初的8个月内收集犬粪样112份,其中南京地区家庭单养的腹泻犬粪便43份,某养犬场群养健康犬粪便30份,沈阳地区某养犬基地群养健康犬粪便39份,用套式PCR方法检测犬冠状病毒(CCV).结果显示,南京家庭单养腹泻病犬CCV阳性率为40.9%(18/43),CCV检出率与季节相关,冬季的检出率较高.健康犬阳性率为84.1%(58/69),其中沈阳某场健康犬CCV的感染率(87.2%)高于南京某犬场(80.0%).取南京腹泻犬和沈阳健康犬阳性样本各2份测序,结果表明,4个样本M基因212bp的序列与GenBank登录的中国大熊猫源的CCV同源性最高(94.8～96.7),并且南京和沈阳CCV毒株之间存在一定序列差异.所有阳性样本用CCV基因型鉴别PCR鉴定,均为CCVⅡ型.

faecal samples of dogs collected from August 2003~January 2004 were tested for the presence of Canine coronavirus (CCV) using a nested-PCR with conserved primers for the M gene.18 out of 43 diarrhea feces from dogs housed singly of Nanjing city were CCV positively, and the CCV detectable rate were positively correlated with seasons, it was higher in winter than that of in summer and autumn. 58 out of 69 feces from heathy dogs trained in groups were positive for CCV, and the positive ratio of samples of dog schools from Shenyang (34/39) was higher than that of from Nanjing(24/30). Sequence analysis of 4 CCV M gene fragments, two from positive diarrhea samples of Nanjing and two from heathy samples of Shenyang, showed the presence of the CCVs with high similarity (94.8%~96.7%) to the CCV of giant panda from China in the GenBank. All the CCVs detected in this survey were type Ⅱ confirmed by a PCR gene typing.