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通过免疫磁珠PCR快速检测艾滋病患者粪便样本中的鸟分枝杆菌

Rapid detection of Mycobacterium avium in stool samples from AIDS patients by immunomagnetic PCR.

作者信息

Li Z, Bai G H, von Reyn C F, Marino P, Brennan M J, Gine N, Morris S L

机构信息

Division of Bacterial Products, Food and Drug Administration, Bethesda, Maryland 20892, USA.

出版信息

J Clin Microbiol. 1996 Aug;34(8):1903-7. doi: 10.1128/jcm.34.8.1903-1907.1996.

Abstract

Direct PCR detection of bacteria in clinical samples is often hindered by the presence of compounds that inhibit the PCR. To improve and accelerate the diagnosis of Mycobacterium avium-M. intracellulare complex infections, an immunomagnetic PCR (IM-PCR) assay was developed. This IM-PCR procedure combines the separation of mycobacteria by antimycobacterial monoclonal antibody coupled to magnetic beads with an M. avium-M. intracellulare complex-specific PCR protocol based on 16S rRNA gene sequences. As few as 10 M. avium bacilli were detected in spiked human stool samples, a clinical specimen usually refractory to conventional PCR analysis, by the IM-PCR method. Moreover, M. avium organisms were detected in about 24 h in 18 of 22 culture-confirmed fecal samples from AIDS patients. This IM-PCR protocol should allow for the rapid and sensitive detection of M. avium isolates in clinical specimens.

摘要

临床样本中细菌的直接聚合酶链反应(PCR)检测常常受到抑制PCR的化合物的阻碍。为了改进并加速鸟分枝杆菌-胞内分枝杆菌复合群感染的诊断,研发了一种免疫磁珠PCR(IM-PCR)检测方法。该IM-PCR程序将与磁珠偶联的抗分枝杆菌单克隆抗体对分枝杆菌的分离,与基于16S rRNA基因序列的鸟分枝杆菌-胞内分枝杆菌复合群特异性PCR方案相结合。通过IM-PCR方法,在加标的人类粪便样本(一种通常难以用传统PCR分析的临床标本)中,检测到低至10条鸟分枝杆菌。此外,在22份来自艾滋病患者且经培养确认的粪便样本中,有18份在约24小时内检测到鸟分枝杆菌。这种IM-PCR方案应能实现临床标本中鸟分枝杆菌分离株的快速、灵敏检测。

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