Morcillo N, Zumarraga M, Alito A, Dolmann A, Schouls L, Cataldi A, Kremer K, van Soolingen D
Mycobacterial Regional Reference Laboratory, Tuberculosis Control Programme of Buenos Aires Province, Argentina.
Int J Tuberc Lung Dis. 2002 Nov;6(11):959-65.
Cetrangolo Hospital, Vicente Lopez, Argentina, 1995-1999.
To describe a home-made reverse-line blot hybridisation assay for the detection of rifampicin resistance-associated mutations in the rpoB gene of Mycobacterium tuberculosis, and to evaluate the usefulness of this rifampicin oligonucleotide, or 'RIFO' assay, to predict rifampicin resistance.
A total of 135 M. tuberculosis isolates from the Cetrangolo Hospital were tested using the RIFO assay, the proportion method and the Mycobacterial Growth Indicator Tube (MGIT 960). In addition, 120 drug-susceptible isolates from the Netherlands were included.
The results obtained with the proportion method and the MGIT 960 system were in full agreement. In the RIFO assay, 90 of the 97 rifampicin-resistant isolates were correctly identified (sensitivity 92.8%, positive predictive value 100%). All of the drug-susceptible isolates were correctly predicted by the RIFO assay.
With this home-made molecular test, rifampicin resistance in M. tuberculosis can be predicted in colonies isolated in culture in only 1 day, and can therefore shorten the laboratory turn around time for rifampicin susceptibility testing by weeks. In principle the test can also be applied directly to Zichl-Neelsen slides and clinical material, as has been demonstrated for another reverse-line blot-based assay for M. tuberculosis, spoligotyping.
1995 - 1999年，阿根廷比森特·洛佩斯的切特兰戈洛医院。
描述一种用于检测结核分枝杆菌rpoB基因中利福平耐药相关突变的自制反向线杂交检测方法，并评估这种利福平寡核苷酸（“RIFO”）检测方法预测利福平耐药性的实用性。
使用RIFO检测法、比例法和分枝杆菌生长指示管（MGIT 960）对切特兰戈洛医院的135株结核分枝杆菌分离株进行检测。此外，还纳入了来自荷兰的120株药物敏感分离株。
比例法和MGIT 960系统获得的结果完全一致。在RIFO检测中，97株利福平耐药分离株中有90株被正确鉴定（敏感性92.8%，阳性预测值100%）。RIFO检测正确预测了所有药物敏感分离株。
通过这种自制的分子检测方法，仅需1天就能在培养分离出的菌落中预测结核分枝杆菌对利福平的耐药性，因此可以将利福平药敏试验的实验室周转时间缩短数周。原则上，该检测方法也可直接应用于萋尼氏涂片和临床材料，就像另一种基于反向线杂交的结核分枝杆菌检测方法——间隔寡核苷酸分型法所证明的那样。
