聚合酶链反应-寡核苷酸探针检测结核菌耐药基因的临床应用

目的探讨聚合酶链反应（PCR）-寡核苷酸探针检测结核菌耐药基因的临床应用价值。方法耐多药肺结核患者80例分为观察组40例和常规组40例，应用PCR-寡核苷酸探针反向斑点杂交技术测定基因耐药情况和突变类型。结果KatG、inhA、rpoB和embB基因突变率分别为90．9％、84．8％、51．98％和58．1％；寡核苷酸探针膜杂交检测rpoB基因突变的灵敏度为98．0％，特异度为100％，阳性预测值为82．61％，与传统药敏试验的符合率为62．50％；寡核苷酸探针膜杂交检测KatG基因突变的灵敏度为50．56％，特异度为75％，阳性预测值为68．75％，与传统药敏试验的符合率为46．43％；观察组病灶显著吸收、吸收、不变、恶化（32．5％、62．5％、5．0％、0％）均高于常规组（25．O％、50．0％、20．0％、5．0％）（X^2=3．98，X^2=3．92，P〈0．05；X^2=6．78，X^2=6．80，P〈0．01）；观察组治疗后3个月、6个月、9个月痰菌阴转率分别为65．0％、77．5％、85．0％，高于常规组（37．5％、50．0％、60．o％）（X^2=3．86，X^2=3．85，X^2=3．89，均P〈0．05）。结论寡核苷酸探针膜杂交技术一次分析可获得结核菌耐3种药物的4个基因；观察组疗效高于常规组。

Clinical application of mutations drug-resistant tuberculosis by polymerase chain reaction-oligonucleotide

Objective To investigate the clinical application of mutations drug-resistant tuberculosis (MDR-TB) by polymerase chain reaction(PCR)-oligonucleotide probes. Methods 80 cases of MDR-TB were divided into observed group 40 cases and conventional group 40 cases, the resistant gene and the type of mutation were determined by polymerase chain reaction (PCR)-oligonucleotide probes. Results 78 isolates of 4 kinds of resistance genes (KatG,inhA,rpoB,embB) ,the mutation rates were 90.9% ,84.8% ,51.98% and 58.1% ;the gene of rpoB sensitivi-ty of 98.0% and specificity of 100% ,positive predictive value of 82.61% ,the rate of according with traditional drug susceptibility testing were 62.50% by polymerase chain reaction (PCR)-oligonucleotide probes;the serisitivity of the KatG gene mutation is of 50.56% ,specificity of 75% ,positive predictive value of 68.75% ,the rate of according with traditional drug susceptibility testing were 46.43% by polymerase chain reaction (PCR)-oligonucleotide probes;the observed group in focus significant absorption, absorption, invariability, deterioration were 32.5% ,62.5% ,5.0%, 0% ,higher than the conventional group(25.0% ,50.0% ,20.0% ,5.0% ) (χ2=3.98,χ2=3.92, P<0.05;χ2=6.78,χ2=6.80,P<0.01) ;observed group in 3 months after treatment,6 months,9 months sputum the conversion negative rate in observed group (65.0%, 77.5%, 85.0% ) higher than the conventional group (37.5%, 50.0%, 60.0%) (χ2=3.86,χ2=3.85,χ2=3.89,P both<0.05). Conclusion An analysis available 3 kind of MDR-TB, 4 kind gene by PCR-oligonucleotide;the observed group in curative effect is better than the conventional group in pa-tients with MDR-TB.