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利用细胞培养、细胞培养-聚合酶链反应联用技术及直接聚合酶链反应检测韩国牡蛎中的人腺病毒和肠道病毒。

Detection of human adenoviruses and enteroviruses in Korean oysters using cell culture, integrated cell culture-PCR, and direct PCR.

作者信息

Choo Yoe-Jin, Kim Sang-Jong

机构信息

School of Biological Sciences, College of Natural Sciences, Seoul National University, Seoul 151-747, Republic of Korea.

出版信息

J Microbiol. 2006 Apr;44(2):162-70.

Abstract

Oysters are known to be carriers of food-born diseases, but research on viruses in Korean oysters is scarce despite its importance for public health. We therefore tested oysters cultivated in Goheung, Seosan, Chungmu, and Tongyeong, for viral contamination using cell culture and integrated cell culture PCR (ICC-PCR) with Buffalo green monkey kidney (BGMK) and human lung epithelial (A549) cells. Additional screens via PCR, amplifying viral nucleic acids extracted from oysters supplemented our analysis. Our methods found 23.6%, 50.9%, and 89.1% of all oysters to be positive for adenoviruses when cell culture, ICC-PCR, and direct PCR, respectively, was used to conduct the screen. The same methodology identified enteroviruses in 5.45%, 30.9%, and 10.9% of all cases. Most of the detected enteroviruses (81.3%) were similar to poliovirus type 1; the remainder resembled coxsackievirus type A1. A homology search with the adenoviral sequences revealed similarities to adenovirus subgenera C (type 2, 5, and 6), D (type 44), and F (enteric type 40 and 41). Adenovirus-positive samples were more abundant in A549 cells (47.3%) than in BGMK cells (18.2%), while the reverse was true for enteroviruses (21.8% vs. 14.5%). Our data demonstrate that Korean oysters are heavily contaminated with enteric viruses, which is readily detectable via ICC-PCR using a combination of A549 and BGMK cells.

摘要

已知牡蛎是食源性疾病的载体,但尽管对公众健康很重要,韩国牡蛎中病毒的研究却很匮乏。因此,我们使用细胞培养和整合细胞培养PCR(ICC-PCR),分别用非洲绿猴肾(BGMK)细胞和人肺上皮(A549)细胞,对在高兴、瑞山、忠武和统营养殖的牡蛎进行了病毒污染检测。通过PCR对从牡蛎中提取的病毒核酸进行扩增的额外筛查补充了我们的分析。当分别使用细胞培养、ICC-PCR和直接PCR进行筛查时,我们的方法发现所有牡蛎中腺病毒阳性率分别为23.6%、50.9%和89.1%。相同的方法在所有病例中分别检测到5.45%、30.9%和10.9%的肠道病毒。检测到的大多数肠道病毒(81.3%)与1型脊髓灰质炎病毒相似;其余的与A1型柯萨奇病毒相似。对腺病毒序列的同源性搜索显示与腺病毒亚属C(2型、5型和6型)、D(44型)和F(肠道40型和41型)相似。腺病毒阳性样本在A549细胞中(47.3%)比在BGMK细胞中(18.2%)更丰富,而肠道病毒则相反(21.8%对14.5%)。我们的数据表明,韩国牡蛎被肠道病毒严重污染,通过使用A549和BGMK细胞组合的ICC-PCR很容易检测到。

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