Javed M Tariq, Aranaz Alicia, de Juan Lucía, Bezos Javier, Romero Beatriz, Alvarez Julio, Lozano Cristina, Mateos Ana, Domínguez Lucas
Grupo de Vigilancia Sanitaria Veterinaria, Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Tuberculosis (Edinb). 2007 Sep;87(5):437-45. doi: 10.1016/j.tube.2007.04.002. Epub 2007 Jun 13.
Spoligotyping is a typing tool used worldwide for epidemiological studies on Mycobacterium tuberculosis complex organisms; however it has received little attention regarding improvement of its discriminatory power (DP). In this study, we have evaluated a spoligotyping membrane prepared with 25 novel spacer sequences selected from a previous study [van der Zanden AG, Kremer K, Schouls LM. Improvement of differentiation and interpretability of spoligotyping for Mycobacterium tuberculosis complex isolates by introduction of new spacer oligonucleotides. J Clin Microbiol 2002;40:4628-39] on 308 M. bovis and 88 M. caprae Spanish isolates in comparison with the traditional spoligotyping membrane. The results obtained by combining the two membranes together revealed an improvement of 45 patterns instead of 31. The spacers used in the second membrane were able to distinguish 8 out of the 16 M. bovis types that had more than one isolate. Seven of these types were differentiated into two subtypes with the second-generation membrane, while spb-7, the most prevalent in Spain, was further differentiated into eight subtypes. This second-generation membrane also differentiates M. bovis from M. caprae. A set of 39 spacers (1, 2, 4-8, 10-15, 17-21, 23, 26-32, 37, 44-49, 51-54, 56 and 57) contain all the DP for both M. bovis and M. caprae isolates; and a set of 35 spacers (1, 2, 4-8, 10-15, 17-21, 26-32, 37, 44-48, 52-54 and 57) had all the DP for the M. bovis isolates. Our results show that the research on new spacers and the design of a new membrane may be useful for epidemiological studies of M. bovis and M. caprae isolates.
间隔寡核苷酸分型法是一种在全球范围内用于结核分枝杆菌复合群生物体流行病学研究的分型工具；然而，关于提高其鉴别能力（DP），该方法很少受到关注。在本研究中，我们评估了一种间隔寡核苷酸分型膜，该膜由从先前研究中[范德赞登AG，克雷默K，舒尔斯LM。通过引入新的间隔寡核苷酸提高结核分枝杆菌复合群分离株间隔寡核苷酸分型法的鉴别能力和可解释性。《临床微生物学杂志》2002年；40：4628 - 39]选择的25个新间隔序列制备而成，用于对308株牛分枝杆菌和88株西班牙山羊分枝杆菌分离株进行检测，并与传统的间隔寡核苷酸分型膜进行比较。将两种膜组合在一起得到的结果显示，可区分的模式从31种增加到了45种。用于第二种膜的间隔序列能够区分16种牛分枝杆菌类型中超过1株的8种类型。其中7种类型通过第二代膜进一步分为两个亚型，而在西班牙最为常见的spb - 7型进一步分为8个亚型。这种第二代膜也能区分牛分枝杆菌和山羊分枝杆菌。一组39个间隔序列（1、2、4 - 8、10 - 15、17 - 21、23、26 - 32、37、44 - 49、51 - 54、56和57）包含了区分牛分枝杆菌和山羊分枝杆菌分离株的所有鉴别能力；一组35个间隔序列（1、2、4 - 8、10 - 15、17 - 21、26 - 32、37、44 - 48、52 - 54和57）具有区分牛分枝杆菌分离株的所有鉴别能力。我们的结果表明，对新间隔序列的研究和新膜的设计可能有助于牛分枝杆菌和山羊分枝杆菌分离株的流行病学研究。
