Molecular methods for Mycobacterium tuberculosis strain typing: a users guide.
There are now a wide range of techniques available to type Mycobacterium tuberculosis, the problem is to choose the correct technique. For large scale epidemiological studies the portability and standardization of IS6110 restriction fragment length polymorphism (RFLP) means that this remains the gold standard technique. In the next few years the internationally standard mycobacterial interspersed repetitive unit (MIRU) may come to challenge this primacy. Low copy number stains remain a problem and these can be typed by either polymorphic Guanine cytosine-rich repetitive sequence (PGRS) or MIRU-variable numbers of tandem repeat (VNTR). To confirm whether strains are part of a true cluster PGRS remains the method of choice. For local outbreaks and investigations of laboratory cross contamination where speed is of greatest importance suspect strains should be initially investigated using a PCR-based method. The superior reproducibility and discrimination of MIRU-VNTR means that these methods should be favoured. If matches are found, then further confirmation of identity can be achieved using IS6110 RFLP or PGRS if the strains prove to have a low IS6110 copy number.
Kanduma
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Clinical Laboratory, Kilimanjaro Christian Medical College, PO Box 3010, Moshi, Tanzania.
McHugh
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Gillespie
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D015373:Bacterial Typing Techniques / Q000379:methods*; D005838:Genotype; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D014376:Tuberculosis / Q000453:epidemiology / Q000382:microbiology