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通过活菌计数技术测定幽门螺杆菌的预形成脲酶活性——临床意义。

Pre-formed urease activity of Helicobacter pylori as determined by a viable cell count technique--clinical implications.

作者信息

Xia H X, Keane C T, O'Morain C A

机构信息

Department of Clinical Microbiology, St. James's Hospital, Dublin, Ireland.

出版信息

J Med Microbiol. 1994 Jun;40(6):435-9. doi: 10.1099/00222615-40-6-435.

Abstract

The pre-formed urease activity of three NCTC reference strains and five clinical isolates of Helicobacter pylori was determined at room temperature (21 degrees C) and 37 degrees C by a viable cell count technique with a conventional urea slope test (Christensen's agar) as well as the commercial CLO-test. The urease activity of two gastroduodenal commensals, Proteus mirabilis and Klebsiella pneumoniae, was also tested. H. pylori strains produced positive reactions with viable cell counts of 10(6)-10(8) cfu within 30 min and with counts of 10(3)-10(6) cfu within 2 h. For some strains, smaller numbers of organisms were needed with the CLO-test than with the conventional test, and incubation of the CLO-test strips at 37 degrees C slightly decreased the number of organisms required for positive results. P. mirabilis produced a positive result on urea slopes with an initial inoculum of 10(7)-10(8) cfu at 2 h, but no positive reaction occurred for K. pneumoniae at 12 h, even with an initial inoculum of 10(11) cfu. However, both P. mirabilis and K. pneumoniae gave a positive result after incubation for 24 h with initial inocula of < 10(1) cfu and 10(3)-10(4) cfu respectively. Incubation at 37 degrees C significantly reduced the inoculum size of these organisms required for a positive result after incubation for 4 h when tested with the slopes, but not with the CLO-test. These findings indicate that H. pylori possesses much greater pre-formed urease activity than P. mirabilis and K. pneumoniae. False negative results for clinical detection of H. pylori in gastroduodenal biopsies may be due to small numbers of organisms, especially after treatment with antimicrobial agents, and false positive results may arise from gastroduodenal commensals or contaminants.

摘要

采用活菌计数技术,通过传统尿素斜面试验(克里斯滕森琼脂)以及商用CLO试验,在室温(21摄氏度)和37摄氏度下测定了3株幽门螺杆菌NCTC参考菌株和5株临床分离株的预形成脲酶活性。还检测了两种胃十二指肠共生菌奇异变形杆菌和肺炎克雷伯菌的脲酶活性。幽门螺杆菌菌株在30分钟内活菌数为10(6)-10(8) cfu时产生阳性反应,在2小时内活菌数为10(3)-10(6) cfu时产生阳性反应。对于一些菌株,CLO试验所需的菌数比传统试验少,并且将CLO试验条在37摄氏度下孵育会略微减少产生阳性结果所需的菌数。奇异变形杆菌在2小时时初始接种量为10(7)-10(8) cfu时在尿素斜面上产生阳性结果,但肺炎克雷伯菌在12小时时即使初始接种量为10(11) cfu也未产生阳性反应。然而,奇异变形杆菌和肺炎克雷伯菌在初始接种量分别<10(1) cfu和10(3)-10(4) cfu孵育24小时后均产生阳性结果。用斜面试验检测时,在37摄氏度下孵育4小时后,这些菌产生阳性结果所需的接种量显著减少,但用CLO试验检测时则不然。这些发现表明,幽门螺杆菌具有比奇异变形杆菌和肺炎克雷伯菌更强的预形成脲酶活性。胃十二指肠活检中幽门螺杆菌临床检测的假阴性结果可能是由于菌数少,尤其是在使用抗菌药物治疗后,而假阳性结果可能源于胃十二指肠共生菌或污染物。

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