DETECTION OF TOXIGENIC CLOSTRIDIUM-DIFFICILE IN STOOL SPECIMENS BY THE POLYMERASE CHAIN-REACTION
DETECTION OF TOXIGENIC CLOSTRIDIUM-DIFFICILE IN STOOL SPECIMENS BY THE POLYMERASE CHAIN-REACTION
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DOI：
10.1093/infdis/167.2.455
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发表时间：
1993-02-01
影响因子：
6.4
通讯作者：
UENO, K
中科院分区：
文献类型：
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作者：
KATO, N;OU, CY;UENO, K
采用聚合酶链反应（PCR）技术扩增毒素a基因片段，直接从抗生素相关性腹泻患者的粪便标本中检测出产毒素艰难梭菌。虽然从粪便标本制备的DNA中识别出了pcr抑制物质，但在苯酚-氯仿萃取后使用离子交换柱消除了抑制物质。最终对39份粪便标本进行PCR评价。产毒艰难梭菌的PCR检测结果与细胞培养实验结果完全一致；12例pcr阳性粪便标本细胞毒素检测均为阳性，27例pcr阴性粪便标本细胞毒素检测均为阴性。从所有pcr阳性标本中培养出产毒艰难梭菌。这些结果提示，PCR扩增可能是艰难梭菌相关性腹泻和结肠炎实验室诊断的有效方法。
Polymerase chain reaction (PCR) amplification of a segment of the toxin A gene was used to detect toxigenic Clostridium difficile directly from stool specimens of patients with antibiotic-associated diarrhea. Although PCR-inhibitory substances were recognized in DNA prepared from stool specimens, the inhibitory substances were eliminated by using an ion-exchange column after phenol-chloroform extraction. Eventually, 39 stool specimens were evaluated by PCR. PCR results for detection of toxigenic C. difficile were in complete agreement with cell culture assay results; all 12 PCR-positive stool specimens were positive by cytotoxin assay, and all 27 PCR-negative specimens were negative by cytotoxin assay. Toxigenic C. difficile was cultured from all PCR-positive specimens. These results suggest that PCR amplification may be an effective method for laboratory diagnosis of C. difficile associated diarrhea and colitis.