薄层凝胶TLA+PNB法培养和鉴定结核杆菌的应用评价

摘要/Abstract

摘要： 目的通过与L-J固体培养，16SrDNA核酸序列测序方法比较，评价一种快速结核分枝杆菌培养和鉴定方法（薄层凝胶TLA+PNB）的应用价值。方法实验样本来源于北京胸科医院疑似结核病例，包括痰样本和气管分泌物。所有的样本用NaOH/NALC方法前处理，然后离心用PBS中和后接种于L-J固体培养基和TLA+PNB固体培养基上。对培养阳性的标本，水煮法提取DNA后PCR扩增16S rDNA核酸序列并测序。最后分别计算2种培养基的敏感度，特异度，培养时间，污染率和TLA+PNB菌种鉴定的敏感度，特异度。结果实验共包含 211份临床样本。对于抗酸染色阳性的标本，培养阳性率为 77.4%,而对于抗酸染色阴性的标本，L-J 和TLA培养阳性率均为 8.2%。TLA 和L-J敏感度均为 84.2%，特异度均为100%。TLA+PNB培养基在 48例培养阳性的标本中发现 8例 NTM，敏感度为88.9%,特异度为100%。污染率TLA 和L-J分别为4.7% 和1.2%。平均报告阳性结果时间 TLA 和L-J分别为10.6-d和22.9-d (P<0.001)。结论 TLA+PNB 培养和鉴定方法是一种快速、可靠、廉价的结核分枝杆菌诊断方法。在基层实验室中，有很好的推广价值。

关键词: 分枝杆菌, 结核, 细菌学技术, 凝胶类, 评价研究

Abstract: Objective evaluate the clinical value of thin-layer agar (TLA+PNB) on the diagnosis of TB and identification of mycobacterial species.  Methods 211 samples including sputum and extra-pulmonary specimens from suspects with a clinical diagnosis of TB were decontaminated using NaOH/NALC, centrifuged, and then stained with Ziehl-Neelsen for the detection of acid-fast bacilli (AFB), cultured on L-J medium and TLA+PNB medium and identified by 16S rDNA sequencing. Sensitivity and specificity, growth detection time and contamination rate were calculated for both media.  Results Of AFB-positive samples, 77.4% were culture positive. Of AFB-negative samples, 8.2% were both L-J and TLA culture positive. The sensitivity and specificity of both L-J and TLA culture were 84.2% and 100%, respectively. 8 non-tuberculosis mycobacteria of 48 cuture-positive samples were detected by TLA+PNB, in which the sensitivity and specificity were 88.9% and 100%, respectively. Contamination rates were 4.7% for TLA and 1.2% for L-J. Median time of a positive culture was 10.6 days for TLA and 22.9 days for L-J (P<0.001).  Conclusions The TLA+PNB method is an inexpensive, rapid and reliable alternative for detectingM. tuberculosis .

Key words: Mycobactorium tuberculosis, baeteriological techniques, gels, evaluation studies

中图分类号:

R446.5

戴广明, 夏辉, 马异峰, 刘冠, 李云絮, 尚媛媛, 付育红, 赵雁林. 薄层凝胶TLA+PNB法培养和鉴定结核杆菌的应用评价[J]. 中国防痨杂志, 2010, 32(9): 109-112.

Dai Guangming,Xia Hui,Ma Yifeng,Liu Guan,Li Yunxv,Shang Yuanyuan,Fu Yuhong,Zhao Yanlin. Evaluation of a method for the rapid culture and identification of mycobacterial species-TLA+PNB[J]. Chinese Journal of Antituberculosis, 2010, 32(9): 109-112.

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