摘要
目的：结核分枝杆菌散在分布重复单位及可变数目串联重复序列（ MIRU-VNTR）是一项应用于结核分枝杆菌分子分型的技术，本研究基于该技术旨在建立一种直接应用临床痰液标本检测结核分枝杆菌的实验室方法。方法：根据扩增条件及临床检测的实际需要，优化选择八个 MIRU-VNTR 位点（ MTUB21、MTUB04、QUB18、QUB26、QUB11b、MIRU31、MIRU10和MIRU26）作为检测靶标，收集130例肺结核患者和200例非肺结核患者（其他肺部疾病）的痰液样本，分别以传统的罗氏培养法和临床诊断的方法为标准，用MIRU-VNTR分析和荧光定量聚合酶链反应（ FQ-PCR ）对上述样本进行检测。结果：以罗氏培养法为标准时， MIRU-VNTR 的灵敏度为93.1％，特异度为97.7％；FQ-PCR 检测的灵敏度为94.0％，特异度为96.7％，MIRU-VNTR分析与FQ-PCR检测结果准确度差异无统计学意义（χ2＝0.352， P＝0.569）。以临床诊断为标准时， MIRU-VNTR分析的灵敏度为86.9％，特异度为100％；FQ-PCR 检测的灵敏度为87.7％，特异度为99.0％，两种方法检测结果准确度差异无统计学意义（χ2＝0.030， P＝0.862），且MIRU-VNTR分析中未发现假阳性结果。在MIRU-VNTR检测为阳性的结果中，98.2％（111／113）的样本分子编码互不相同，只有1.8％（2／113）分子编码一致（皆为54685538）。结论：应用临床痰液标本建立的基于MIRU-VNTR的结核分枝杆菌快速检测方法有着较好的应用价值。
Objective： To establish a laboratory method for detection of Mycobacterium tuberculosis in sputum based on variable number tandem repeat （ VNTR） .Methods： Mycobacterium tuberculosis was tested by VNTR and fluorescent quantitative reverse transcription polymerase chain reaction （ FQ-PCR ） in 130 sputum samples from patients with pulmonary tuberculosis and 200 specimens from patients with other lung diseases.According to the amplification conditions and clinical detection needs, MTUB21, MUTB04, QUB18, QUB26, QUB11b, MIRU31, MIRU10 and MIRU26 were selected as test targets.The results of VNTR and FQ-PCR were compared with Lowenstein-Jensen culture and clinical diagnosis, and analyzed by chi-square test. Results：With the results of L-J culture as the standard, the sensitivity and specificity of VNTR were 93.1%（108/116） and 97.7%（209/214）, and those of FQ-PCR were 94 .0% （ 109/116 ） and 96 .7% （ 207/214 ） , respectively; no significant difference was observed between two groups （χ2 =0.352, P =0.569 ）.Using the clinical diagnosis as the standard, the sensitivity and specificity of VNTR were 86.9% （113/130） and 100% （200/200）, and those of FQ-PCR were 87.7% （114/130） and 99.0%（198/200）, respectively;the difference was not statistically significant （χ2 =0.030, P=0.862）.In 113 VNTR positive samples, the molecular codes differed from each other in 98.2%samples （111/113）;only 2 samples had identical code （5 4 6 8 5 5 3 8 ） .Conclusion：The study suggests that VNTR provides a promising method for diagnosis of clinical tuberculosis.
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2016年第1期61-67,共7页
Journal of Zhejiang University(Medical Sciences)
基金
国家自然科学基金
关键词
分枝杆菌
结核/分离和提纯
结核
肺/诊断
痰液/微生物学
分枝杆菌
结核/遗传学
基因型
串联重复序列
Mycobacterium tuberculosis/isolation ＆amp
purification
Tuberculosis,pulmonary/diagnosis
Sputum/microbiology
Mycobacterium tuberculosis/genetics
Genotype
Tandem repeat sequences
