Suresh Naga, Arora Jyoti, Pant Hema, Rana Tanu, Singh Urvashi Balbir
Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India.
J Microbiol Methods. 2007 Feb;68(2):291-5. doi: 10.1016/j.mimet.2006.09.001. Epub 2006 Oct 27.
Spoligotyping was applied to old (5-11 years) Ziehl-Neelsen (ZN)-stained smears for strain identification and differentiation and to predict the utility of the technique in epidemiological studies. Among 57 DNA samples extracted from ZN slides lying stored at room temperature, 93% (53) amplification was achieved for mpt64 gene. Spoligopatterns were generated from 77.7% (41/53) DNA samples, whereas negative controls did not yield any spoligopatterns. All slides with 2+ (n=20) and 3+ (n=13) positivity while 42% (11/26) of slides with low positivity (< or 1+) showed a good signal and a reproducible pattern. This technique may have application in identification of spoligotypes in control programme implemented areas remote from research laboratory and would also increase our knowledge about the clonal structure of Mycobacterium tuberculosis in the population, when applied to old samples in different locations.
spoligotyping技术应用于旧的（5 - 11岁）齐-尼氏（ZN）染色涂片，用于菌株鉴定和区分，并预测该技术在流行病学研究中的效用。在从室温保存的ZN载玻片上提取的57个DNA样本中，mpt64基因的扩增成功率为93%（53个）。77.7%（41/53）的DNA样本产生了spoligotype图谱，而阴性对照未产生任何spoligotype图谱。所有2 +（n = 20）和3 +（n = 13）阳性的载玻片以及42%（11/26）低阳性（<或1 +）的载玻片显示出良好的信号和可重复的图谱。该技术可能在远离研究实验室的控制项目实施地区的spoligotype鉴定中具有应用价值，并且当应用于不同地点的旧样本时，也将增加我们对人群中结核分枝杆菌克隆结构的了解。
