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聚合酶链反应在分枝杆菌培养物中对结核病的早期诊断的应用。

The use of polymerase chain reaction for early diagnosis of tuberculosis in Mycobacterium tuberculosis culture.

机构信息

Departamento de Análises Clínicas, Universidade Federal de Santa Catarina, Florianópolis, SC, Brasil.

出版信息

Braz J Med Biol Res. 2010 Jun;43(6):543-8. doi: 10.1590/s0100-879x2010007500031. Epub 2010 Apr 23.

Abstract

Early diagnosis plays a vital role in controlling tuberculosis. The conventional methodology is slow, with results taking several weeks, in addition to having low sensitivity, especially in clinical paucibacillary samples. The objective of this study was to evaluate the use of polymerase chain reaction (PCR) on solid medium culture for a rapid diagnosis of tuberculosis, mainly in cases of negative sputum smears. Forty sputum samples were collected from inpatients with tuberculosis treated for less than 2 days. Bacilloscopy, PCR for sputum, culture on Löwestein-Jensen (LJ) solid medium, and daily PCR from culture were performed on each sample. DNA extracted from the BCG vaccine, which contains attenuated bacillus Calmette-Guérin, was used as the positive control. Smear microscopy showed 68.6% sensitivity, 80% specificity, 96% positive predictive value, and 26.7% negative predictive value, with culture on LJ medium as the gold standard. Culture at day 28 showed 74.3% sensitivity and 100% specificity. PCR of DNA extracted from sputum amplified a 1027-bp fragment of the 16s RNA gene, showing 22.9% sensitivity and 60% specificity. PCR performed with DNA extracted from daily culture showed that, from the 17th to the 40th day, the sensitivity (85.7%) and specificity (60%) were constant. We conclude that a 17-day culture is a good choice for rapid diagnosis and to interfere with the transmission chain of tuberculosis.

摘要

早期诊断对于控制结核病至关重要。传统方法速度较慢,结果需要数周时间,并且敏感性较低,尤其是在临床少菌样本中。本研究旨在评估聚合酶链反应(PCR)在固体培养基培养物中的应用,以快速诊断结核病,特别是在痰涂片阴性的情况下。从接受少于 2 天治疗的结核病住院患者中收集了 40 份痰液样本。对每个样本进行了巴氏涂片镜检、痰液 PCR、Löwestein-Jensen(LJ)固体培养基培养和培养物的每日 PCR。将含有减毒卡介苗的 BCG 疫苗的 DNA 作为阳性对照。涂片镜检显示出 68.6%的敏感性、80%的特异性、96%的阳性预测值和 26.7%的阴性预测值,以 LJ 培养基培养作为金标准。第 28 天的培养显示出 74.3%的敏感性和 100%的特异性。从痰液中提取的 DNA 的 PCR 扩增了 16s RNA 基因的 1027bp 片段,显示出 22.9%的敏感性和 60%的特异性。用从每日培养物中提取的 DNA 进行的 PCR 显示,从第 17 天到第 40 天,敏感性(85.7%)和特异性(60%)保持不变。我们得出结论,17 天的培养是快速诊断和干预结核病传播链的良好选择。

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