Le Guyader F, Neill F H, Estes M K, Monroe S S, Ando T, Atmar R L
Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 77030, USA.
Appl Environ Microbiol. 1996 Nov;62(11):4268-72. doi: 10.1128/aem.62.11.4268-4272.1996.
Outbreaks of shellfish-transmitted viral disease occur periodically, but frequently the causative agent is not identified. In November 1993, during investigation of a multistate outbreak of acute gastroenteritis, incriminated lots of oysters were collected. Oyster tissues (stomachs and digestive diverticula) were processed for virus extraction and nucleic acid purification. Human calicivirus sequences were sought by reverse transcriptase PCR using different primer sets. Amplicons were obtained from 9 of 10 shellfish samples from four different lots when primers specific for the outbreak virus strain were used. The specificity of the amplification was confirmed by hybridization. The amplicons from the nine positive oysters were cloned and sequenced. The sequence of each of the clones was identical to the others but showed some variation (7 of 81 bp) from the sequences obtained from the stools of three persons made III by the outbreak.
贝类传播的病毒性疾病暴发时有发生,但致病原往往难以确定。1993年11月,在对一次多州急性肠胃炎暴发进行调查期间,采集了被认定有问题的牡蛎批次。对牡蛎组织(胃和消化盲囊)进行处理以提取病毒并纯化核酸。使用不同引物组通过逆转录聚合酶链反应寻找人杯状病毒序列。当使用针对暴发病毒株的特异性引物时,从四个不同批次的10个贝类样本中的9个获得了扩增子。通过杂交确认了扩增的特异性。对来自9只阳性牡蛎的扩增子进行克隆和测序。每个克隆的序列彼此相同,但与暴发导致三名患者发病的粪便中获得的序列相比有一些差异(81个碱基对中有7个)。