摘要
目的调查北京地区市售牡蛎轮状病毒的污染状况。方法2014年2月至2015年3月，在北京市某大型水产品批发市场采集，每摊位采5只为一组，共56组280只牡蛎。用3种方法对样品前处理：直接处理法、PEG8000沉淀法和蛋白酶K消化．PEG8000沉淀法。然后利用荧光RT-PCR检测样品的A组轮状病毒核酸，采用半巢式反转录．聚合酶链反应方法对A组轮状病毒核酸阳性进行G、P基因分型。对阳性毒株的VP7和VP4基因进行扩增和测序，采用Mega6．06软件构建进化树，建树方法为最大似然法。结果直接处理法、PEG8000沉淀法和蛋白酶K消化．PEG8000沉淀法3种前处理方法A组轮状病毒检出率分别为3．57％（2／56）、7．14％（4／56）和5．38％（3／56）。56组样品中8组A组轮状病毒核酸检测阳性，检出率为14．29％，均在秋冬季检出。轮状病毒基因型组合型为G9／P[8]（2株）和G9／P[N]（6株）。3株轮状病毒VP7基因和1株轮状病毒VP4基因成功测序，进化分析显示与近年来我国流行的G9／P[8]型毒株同源性最高。结论北京地区秋冬季市售部分牡蛎存在轮状病毒污染，有引起食源性疾病的风险。
Objective To investigate the rotavirus contamination in commercial oysters in Beijing. Methods Between February 2014 and March 2015, a total of 280 oysters were collected in a large aquatic market in Beijing, and 5 oysters per stall were collected and classified as one sample. The samples were processed using three kinds of methods ： direct treatment, PEG （ polyethylene glycol） 8 000 precipitation and proteinase K digestion-PEG 8 000 precipitation. Group A rotaviruses were detected by real time RT-PCR, and G/P genotyping was performed using the semi-nested RT-PCR. The VP7 and VP4 genes of positive samples were amplified, sequenced, and the phylogenetic tree was constructed using the maximum likelihood method with MEGA software （version 6. 06）. Results The detection rates of group A rotaviruses of three methods （direct treatment, PEG 8 000 precipitation and proteinase K digestion-PEG 8 000 precipitation） were 3.57% （2/56）, 7.14% （4/56） and 5.38% （3/56）, respectively. Rotaviruses were detected in 8 oysters samples （14. 29% ）, which were collected during autumn and winter. The G/P genotype combination included G9/P[8] （2 strains） and G9/P[N] （6 strains）. The VP7 genes of 3 strains and the VP4 gene of one strain were sequenced successfully, and the phylogenetic analyses demonstrated that these strains had the highest similarity to those G9/P[ 8 ] strains prevailing in recent years in China. Conclusion Group A rotaviruses were detected in some commercial oysters during autumn and winter in Beijing, indicating a risk of foodborne illness.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
2016年第1期41-44,共4页
Chinese Journal of Experimental and Clinical Virology
基金
北京市自然科学基金项目（7132045）
