Al-Quds Nutrition and Health Research Institute, Al-Quds University, Abu-Deis, West Bank, Palestine.
Trop Med Int Health. 2011 Mar;16(3):360-7. doi: 10.1111/j.1365-3156.2010.02697.x. Epub 2010 Dec 16.
To compare the effectiveness and feasibility of an insertion sequence (IS6110)-based polymerase chain reaction (PCR) assay with conventional methods of detecting Mycobacterium tuberculosis and to analyse mutations present in the hot spot region of the RNA polymerase B subunit (rpoB) gene associated with rifampin resistance by DNA sequencing.
Ninety-five sputum samples from 84 clinically suspected cases of tuberculosis were tested for mycobacterial infections by Ziehl Neelsen smear examination, Lowenstein-Jensen culture and IS6110-based PCR assay.
Sensitivity and specificity of the PCR were 94%; the sensitivity of culture was 65%, and of smear tests, 59%. Both smear microscopy and culture had 100% specificity. DNA sequencing data of the 305-bp fragment of the rpoB gene for nine clinical isolates revealed one point mutation at position I572F and double mutations at position S531F in two isolates obtained from two patients who did not respond to the anti-tuberculosis therapy.
IS6110-based PCR can be used routinely in clinical laboratories for rapid detection of Mycobacterium tuberculosis and thus allow early diagnosis and treatment of any contacts by the cheapest method currently available in the Palestinian Authority region. Rapid detection of rifampin resistance isolates will enable efficient treatment of patients and assist in eradication of the disease in the Palestinian territories.
比较插入序列（IS6110）-聚合酶链反应（PCR）检测与传统方法检测结核分枝杆菌的效果和可行性，并通过 DNA 测序分析与利福平耐药相关的 RNA 聚合酶 B 亚单位（rpoB）基因热点区的突变。
对 84 例临床疑似结核病例的 95 份痰标本进行了齐氏染色镜检、Lowenstein-Jensen 培养和基于 IS6110 的 PCR 检测，以检测分枝杆菌感染。
PCR 的敏感性和特异性分别为 94%；培养的敏感性为 65%，涂片检查的敏感性为 59%。涂片镜检和培养均具有 100%的特异性。对来自两名未对抗结核治疗产生反应的患者的 9 株临床分离株 rpoB 基因 305bp 片段的 DNA 测序数据显示，在 2 个分离株中发现 1 个位置 I572F 的点突变和 2 个位置 S531F 的双突变。
基于 IS6110 的 PCR 可常规用于临床实验室，快速检测结核分枝杆菌，从而能够以目前在巴勒斯坦权力机构地区最便宜的方法尽早诊断和治疗任何接触者。快速检测利福平耐药分离株将有助于对患者进行有效的治疗，并有助于在巴勒斯坦领土消灭该疾病。
