PCR-based method to differentiate the subspecies of the Mycobacterium tuberculosis complex on the basis of genomic deletions

被引:216
作者
Huard, RC
Lazzarini, LCD
Butler, WR
van Soolingen, D
Ho, JL
机构
[1] Cornell Univ, Joan & Sanford I Weill Med Coll, Dept Med, Div Int Med & Infect Dis, New York, NY 10021 USA
[2] Cornell Univ, Grad Sch Med Sci, New York, NY 10021 USA
[3] Ctr Dis Control & Prevent, Div AIDS STD & TB Lab Res, Natl Ctr Infect Dis, Atlanta, GA USA
[4] Natl Inst Publ Hlth & Environm, NL-3720 BA Bilthoven, Netherlands
关键词
D O I
10.1128/JCM.41.4.1637-1650.2003
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The classical Mycobacterium tuberculosis complex (MtbC) subspecies include Mycobacterium tuberculosis, Mycobacterium africanum (subtypes I and II), Mycobacterium bovis (along with the attenuated M. bovis bacillus Calmette-Guerin [BCG]), and Mycobacterium microti; increasingly recognized MtbC groupings include Mycobacterium bovis subsp. caprae and "Mycobacterium tuberculosis subsp. canettii" Previous investigations have documented each MtbC subspecies as a source of animal and/or human tuberculosis. However, study of these organisms is hindered by the lack of a single protocol that quickly and easily differentiates all of the MtbC groupings. Towards this end we have developed a rapid, simple, and reliable PCR-based MtbC typing method that makes use of MtbC chromosomal region-of-difference deletion loci. Here, seven primer pairs (which amplify within the loci 16S rRNA, Rv0577, IS1561', Rv1510, Rv1970, Rv3877/8, and Rv3120) were run in separate but simultaneous reactions. Each primer pair either specifically amplified a DNA fragment of a unique size or failed, depending upon the source mycobacterial DNA. The pattern of amplification products from all of the reactions, visualized by agarose gel electrophoresis, allowed immediate identification either as MtbC composed of M. tuberculosis (or M. africanum subtype II), M. africanum subtype I, M. bovis, M. bovis BCG, M. caprae, M. micron, or "M. canettii" or as a Mycobacterium other than MtbC (MOTT). This MtbC PCR typing panel provides an advanced approach to determine the subspecies of MtbC isolates and to differentiate them from clinically important MOTT species. It has proven beneficial in the management of Mycobacterium collections and may be applied for practical clinical and epidemiological use.
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收藏
页码:1637 / 1650
页数:14
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