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采用聚合酶链反应技术从泰国腹泻患者粪便样本中检测艰难梭菌毒素A和B基因。

Detection of Clostridium difficile toxin A and B genes from stool samples of Thai diarrheal patients by polymerase chain reaction technique.

作者信息

Wongwanich Siripan, Rugdeekha Siriporn, Pongpech Pintip, Dhiraputra Chertsak

机构信息

National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi 11000, Thailand.

出版信息

J Med Assoc Thai. 2003 Oct;86(10):970-5.

PMID:14650710
Abstract

The prevalence of Clostridium difficile isolated from stools of Thai adult patients with suspected antibiotic-associated diarrhea (AAD) was 18.64 per cent. The recovery rate of toxin genes (tcdA and tcdB) by polymerase chain reaction (PCR) from stool samples yielded almost the same compared to the recovery rate of the toxin detection by enzyme immunoassay (EIA), which were 44.9 per cent and 46.7 per cent, respectively. Correlation of toxin gene detection by PCR and toxin detection by EIA was 90.6 per cent. All but one stool sample, the tcdA gene was detected together with the tcdB gene. Both genes were always detected together from tox gene-positive strains. Although, there were some discrepancy results for certain samples, the direct PCR-based-detection of C. difficile tox genes in stool samples seems to be the appropriate method for the diagnosis of C. difficile diarrhea. The PCR assay should be a recommended technique to be used routinely in laboratories. Further optimization of the technique to increase the sensitivity of the PCR assays is still needed. However, a quantitative isolation of the organism from stools of suspected antibiotic-associated diarrhea (AAD) or antibiotic-associated colitis (AAC) patients may give some evidence for clinicians in hospitals who cannot perform PCR-based or EIA-based techniques, since 48.6 per cent of the isolates were demonstrated as toxigenic strains.

摘要

从疑似抗生素相关性腹泻(AAD)的泰国成年患者粪便中分离出艰难梭菌的患病率为18.64%。通过聚合酶链反应(PCR)从粪便样本中检测毒素基因(tcdA和tcdB)的回收率与酶免疫测定(EIA)检测毒素的回收率几乎相同,分别为44.9%和46.7%。PCR检测毒素基因与EIA检测毒素的相关性为90.6%。除一份粪便样本外,所有样本中tcdA基因均与tcdB基因同时被检测到。从毒素基因阳性菌株中总能同时检测到这两个基因。尽管某些样本存在一些差异结果,但基于PCR直接检测粪便样本中艰难梭菌毒素基因似乎是诊断艰难梭菌腹泻的合适方法。PCR检测应成为实验室常规使用的推荐技术。仍需要进一步优化该技术以提高PCR检测的灵敏度。然而,对于无法进行基于PCR或基于EIA技术的医院临床医生而言,从疑似抗生素相关性腹泻(AAD)或抗生素相关性结肠炎(AAC)患者粪便中定量分离该病原体可能会提供一些依据,因为48.6%的分离株被证明是产毒菌株。

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