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通过β-珠蛋白基因扩增检测沙眼衣原体的生殖器样本中,有很大一部分含有少量细胞DNA。

An important proportion of genital samples submitted for Chlamydia trachomatis detection by PCR contain small amounts of cellular DNA as measured by beta-globin gene amplification.

作者信息

Coutlée F, de Ladurantaye M, Tremblay C, Vincelette J, Labrecque L, Roger M

机构信息

Département de Microbiologie et Immunologie, Université de Montréal, Montréal, Québec, Canada.

出版信息

J Clin Microbiol. 2000 Jul;38(7):2512-5. doi: 10.1128/JCM.38.7.2512-2515.2000.

Abstract

We assessed the quality of genital samples submitted for Chlamydia trachomatis detection by PCR by a second PCR assay for the presence of human beta-globin DNA. Endocervical and urethral samples were first tested by the COBAS AMPLICOR C. trachomatis assay (Roche Diagnostic Systems) with an internal control and were then amplified for the presence of beta-globin DNA with primers PC04 and GH20. Samples that contained inhibitors were retested after dilution 1:10. A total of 407 genital samples (311 endocervical swabs from 311 women and 96 urethral swabs from 95 men and 1 woman) collected over a 1-month period were evaluated. The internal control could not be amplified, despite dilution, from 3 of 23 samples that were retested after dilution because of inhibition, leaving 404 samples that could be analyzed by PCR. Eleven samples tested positive for C. trachomatis. Thirty (7.4%) of the 404 samples were negative for beta-globin. Twelve of the 23 undiluted samples that contained inhibitors tested positive for beta-globin DNA. Amplification of beta-globin DNA in samples submitted for C. trachomatis detection by the COBAS AMPLICOR C. trachomatis assay demonstrated that an important proportion of the samples did not contain cellular DNA. Assessment of the quality of the samples for PCR analysis by beta-globin amplification is feasible but cannot replace use of the internal control.

摘要

我们通过针对人β-珠蛋白DNA存在情况的第二次聚合酶链反应(PCR)检测,评估了提交用于通过PCR检测沙眼衣原体的生殖器样本的质量。首先使用带有内部对照的COBAS AMPLICOR沙眼衣原体检测法(罗氏诊断系统公司)对宫颈内膜和尿道样本进行检测,然后使用引物PC04和GH20对样本进行扩增以检测β-珠蛋白DNA的存在。含有抑制剂的样本在1:10稀释后重新检测。对在1个月期间收集的总共407份生殖器样本(311名女性的311份宫颈内膜拭子以及95名男性和1名女性的96份尿道拭子)进行了评估。尽管进行了稀释,但在因抑制作用而重新检测的23份样本中有3份样本的内部对照无法扩增,因此留下404份可通过PCR进行分析的样本。11份样本沙眼衣原体检测呈阳性。404份样本中有30份(7.4%)β-珠蛋白检测呈阴性。23份未稀释的含有抑制剂的样本中有12份β-珠蛋白DNA检测呈阳性。通过COBAS AMPLICOR沙眼衣原体检测法提交用于检测沙眼衣原体的样本中β-珠蛋白DNA的扩增表明,相当一部分样本不含有细胞DNA。通过β-珠蛋白扩增评估用于PCR分析的样本质量是可行的,但不能替代内部对照的使用。

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