【摘要】：目的：應(yīng)用GenoType MTBDRplus方法和傳統(tǒng)比例法,對(duì)結(jié)核分枝桿菌利福平耐藥性進(jìn)行檢測(cè)研究對(duì)比。GenoType MTBDRplus方法采用核酸反向線性探針雜交技術(shù)檢測(cè)與利福平耐藥密切相關(guān)的rpoB基因,可在24小時(shí)內(nèi)完成檢測(cè)。傳統(tǒng)比例法需要3個(gè)月左右完成檢測(cè),雖然方法簡(jiǎn)單、經(jīng)濟(jì)、易于推廣,難于滿足臨床對(duì)結(jié)核病診斷和治療的需求,限制了其臨床的應(yīng)用。GenoType MTBDRplus方法為早期診斷耐藥結(jié)核病提供快速檢測(cè)依據(jù),利于臨床早期合理治療。 方法：標(biāo)本來(lái)自于2011年4月至2011年9月,天津市濱海新區(qū)結(jié)核病防治所就診涂片陽(yáng)性患者的痰標(biāo)本,共收集55例患者的痰標(biāo)本。同一份痰標(biāo)本分別進(jìn)行GenoType MTBDRplus方法和傳統(tǒng)比例法檢測(cè),檢測(cè)結(jié)核分枝桿菌對(duì)利福平耐藥性。GenoType MTBDRplus方法采用核酸反向線性探針雜交技術(shù)檢測(cè)與利福平耐藥密切相關(guān)的rpoB基因,推測(cè)其對(duì)利福平的耐藥性。GenoType MTBDRplus方法檢測(cè)過(guò)程包括,涂陽(yáng)痰標(biāo)本的處理通過(guò)堿處理-中和離心沉淀法,使用GenoLyse試劑盒提取DNA,PCR擴(kuò)增,雜交,結(jié)果判讀完成。傳統(tǒng)比例法檢測(cè)結(jié)核分枝桿菌利福平耐藥性,通過(guò)實(shí)驗(yàn)材料準(zhǔn)備,制備菌懸液和稀釋,接種菌液和培養(yǎng),結(jié)果報(bào)告完成。應(yīng)用Kappa檢驗(yàn)對(duì)兩種方法進(jìn)行一致性的統(tǒng)計(jì)分析。研究在天津市結(jié)核病控制中心參比室進(jìn)行。 結(jié)果：1.55例涂陽(yáng)標(biāo)本中,1例經(jīng)鑒定為非結(jié)核分枝桿菌(NTM),1例培養(yǎng)結(jié)果為污染,1例培養(yǎng)陽(yáng)性而GenoType MTBDRplus無(wú)法判讀�？膳c比例法藥物敏感性試驗(yàn)進(jìn)行對(duì)比的標(biāo)本為52例。2.利福平耐藥性的結(jié)果。以比例法藥敏結(jié)果為判斷標(biāo)準(zhǔn),使用GenoType MTBDRpuls檢測(cè)利福平耐藥性的敏感度為75%,特異度為97.9%,陽(yáng)性預(yù)測(cè)值為75%,陰性預(yù)測(cè)值為97.9%,總一致率為96.2%。對(duì)兩種方法進(jìn)行一致性Kappa檢驗(yàn),K=0.729(P0.01)。 結(jié)論：GenoType MTBDRplus方法檢測(cè)結(jié)核分枝桿菌對(duì)利福平的耐藥性,與傳統(tǒng)比例法檢測(cè)比較其敏感度和特異度,具有較高的一致性。但其較傳統(tǒng)比例法快速,為臨床早期診斷耐藥結(jié)核病提供快速檢測(cè)依據(jù),利于臨床早期合理治療。
[Abstract]:Objective: to detect rifampicin resistance of Mycobacterium tuberculosis by GenoType MTBDRplus and traditional proportional method. Genotype MTBDRplus was used to detect rpoB gene, which is closely related to rifampicin resistance. The test can be completed within 24 hours. The traditional proportional method takes about three months to complete the test, although the method is simple, economical and easy to popularize, and it is difficult to meet the clinical needs for the diagnosis and treatment of tuberculosis. Genotype MTBDRplus provides rapid detection basis for early diagnosis of drug-resistant tuberculosis and is beneficial to early and reasonable clinical treatment. Methods: from April 2011 to September 2011, sputum samples from smear positive patients in Tianjin Binhai New District Tuberculosis Prevention and Control Institute were collected. The resistance of Mycobacterium tuberculosis to rifampicin was detected by GenoType MTBDRplus and traditional proportional method, respectively. Genotype MTBDRplus was used to detect rpoB gene closely related to rifampicin resistance. It was speculated that the resistance of rifampicin to rifampicin was speculated. GenoType MTBDRplus method included the treatment of positive sputum samples by alkali treatment-neutralization centrifugal precipitation method, the extraction and amplification of DNA,PCR with GenoLyse kit, and the results were read out. The rifampicin resistance of Mycobacterium tuberculosis was detected by traditional proportional method. Through the preparation of experimental materials, the suspension and dilution of bacteria were prepared, inoculated and cultured, and the results were reported. Kappa test is used to analyze the consistency of the two methods. The study was carried out in the reference room of Tianjin Tuberculosis Control Center. Results: 1. Among 55 smear positive specimens, 1 case was identified as non-tuberculosis mycobacteria (NTM), 1 case was contaminated, 1 case was positive and GenoType MTBDRplus could not be read. 2. 52 cases were compared with the proportional drug sensitivity test. 2. The result of rifampicin resistance. According to the results of proportional drug sensitivity, the sensitivity, specificity, positive predictive value, negative predictive value and total consistency rate of rifampicin resistance were 75%, 97.9%, 75%, 97.9% and 96.2%, respectively. the sensitivity, specificity, positive predictive value and negative predictive value of rifampicin were 75%, 97.9%, 75% and 97.9%, respectively. The consistency of the two methods was tested by Kappa, K 鈮，

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