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Guide to the Application of Genotyping to Tuberculosis Prevention
and Control
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CDC Tuberculosis Genotyping Laboratory Procedures
IS6110-based RFLP Analysis
TB programs can request the genotyping laboratories to perform
IS6110-based RFLP analysis on isolates that match by both
spoligotyping and MIRU analysis. See Chapter 5, Developing a
Tuberculosis Genotyping Program, for a description of when to
request RFLP analysis. The request for RFLP analysis must list the
cluster number and the specific isolates in that cluster to be typed.
If additional isolates in the cluster are identified at a later
time, the program can request IS6110-based RFLP analysis
of the additional isolates.
Only those isolates listed in the request will be typed by RFLP
analysis. RFLP patterns will be analyzed for matches or non-matches
among the requested isolates. The genotyping laboratory will not
search the entire database for possible RFLP matches outside the
requested cluster. Each isolate typed by RFLP analysis will be assigned
an RFLP pattern designation number. If one of the requested isolates
demonstrates an RFLP pattern that does not match the patterns of
any of the other requested isolates, the isolate will be assigned
a new RFLP pattern designation number, which will be the genotyping
laboratory’s accession number for that isolate plus the band number
for the RFLP pattern. For isolates with matching RFLP patterns,
the lowest accession number in that cluster will be assigned to
all isolates. Again, the RFLP pattern designation number applies
only to isolates within a specific PCR cluster (i.e., the same spoligotype
and MIRU type from the same TB program). Unfortunately, this may
result in two isolates having the same RFLP pattern but different
RFLP pattern designation numbers.
Last Reviewed: 05/18/2008 Content Source: Division of Tuberculosis Elimination
National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention
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