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[利用聚合酶链反应从奶牛中检测产志贺毒素大肠杆菌]

[Detection of verotoxin-producing Escherichia coli using polymerase chain reaction from dairy cattle].

作者信息

Tada H, Itami S, Yamamoto Y, Kobayashi K, Taguchi M, Nakazawa M

机构信息

Tokushima Prefectural Institute of Public Health and Environmental Sciences.

出版信息

Kansenshogaku Zasshi. 1992 Oct;66(10):1383-9. doi: 10.11150/kansenshogakuzasshi1970.66.1383.

Abstract

The vero cytotoxin (VT) is responsible for hemorrhagic colitis and hemolytic uremic syndrome. Polymerase chain reaction (PCR) was used to detect VT-producing coliform bacteria from dairy cattle. It was found that 39 (33.3%) of the 117 fecal samples examined were recognized with VT genes in BGLB enrichment broth by the PCR method (named BGLB-PCR). Of the VT-positive samples, 31 samples (26.5%) were found to have VT-producing Escherichia coli. Frequencies of isolation in younger cattle (under 5 months) were 31.3-32.9%. On the other hand, the PCR method using the bacterial suspension of some colonies from DHL selective isolation medium (named DHL-PCR), was used for 105 samples. The DHL-PCR was validated according to the number of colonies tested for detecting VTEC. When using E. coli strains which have been stored after isolation by the conventional culture method, the VT-producing strains found were 7 (10.3%) of the 68 isolates tested. The 101 out of the 108 VTEC strains from cattle were classified into 14 O groups. 4 O serogroups (O26, O111, O145, O157) from 60% of VTEC positive cattle, were also the most common in humans with diarrhea. All E. coli O157:H7 isolates failed to ferment after 48 hrs and to hydrolyze 4-methyl-umbelliferyl-beta-D-glucuronide (MUG). These results suggests that cattle may play an important role in human VTEC infections. The BGL B-PCR technique is usefull in ecological studies for VT-producing pathogens.

摘要

维罗细胞毒素(VT)可引发出血性结肠炎和溶血尿毒综合征。采用聚合酶链反应(PCR)检测奶牛中产VT的大肠菌群。结果发现,在117份粪便样本中,有39份(33.3%)通过PCR方法(称为BGLB-PCR)在BGLB增菌肉汤中被检测出含有VT基因。在VT阳性样本中,有31份样本(26.5%)被发现含有产VT的大肠杆菌。幼龄牛(5月龄以下)的分离频率为31.3 - 32.9%。另一方面,对105份样本采用了从DHL选择性分离培养基中选取部分菌落的细菌悬液进行PCR的方法(称为DHL-PCR)。根据检测产VTEC的菌落数量对DHL-PCR进行了验证。当使用通过传统培养方法分离后保存的大肠杆菌菌株时,在68株检测菌株中发现产VT菌株有7株(10.3%)。来自牛的108株VTEC菌株中有101株被分为14个O群。来自60% VTEC阳性牛的4个O血清群(O26、O111、O145、O157)在腹泻人群中也最为常见。所有大肠杆菌O157:H7分离株在48小时后均不能发酵,也不能水解4 - 甲基伞形酮基 - β - D - 葡萄糖醛酸苷(MUG)。这些结果表明,牛可能在人类VTEC感染中起重要作用。BGLB-PCR技术在产VT病原体的生态学研究中很有用。

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