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Guide to the Application of Genotyping to Tuberculosis Prevention
and Control
Return to Genotyping Main Menu
CDC Tuberculosis Genotyping Laboratory Procedures
Genotyping Results, Designation of Clusters, and Reporting
Turnaround Times of Genotyping Results
CDC’s contract with the genotyping laboratories requires that the
laboratories genotype all of the eligible isolates by PCR tests
(spoligotyping and MIRU analysis) and report results for at least
90% of isolates to the TB programs within 10 working days of receipt
of the isolates. These rapid turnaround times assume a steady flow
of isolates to the laboratory. Although we expect that these projected
turnaround times will be met most of the time, there are several
reasons for possible delays. The MIRU automated sequencer is limited
to 24 isolates in each run; if a genotyping laboratory receives
large batches at one time, there will be a delay in reporting some
of the results. Technical difficulties (e.g., instrument failure)
may also cause delays.
Under certain circumstances, the TB program can request that the
genotyping laboratories perform IS6110-based RFLP analysis
on isolates that match by both spoligotyping and MIRU analysis.
See “IS6110-based RFLP Analysis” (Chapter 3, CDC Tuberculosis
Genotyping Laboratory Procedures) for details about how the
genotyping laboratory will report RFLP results, and see chapter
5, Developing a Tuberculosis Genotyping Program, for a description
of circumstances where RFLP analysis is appropriate. CDC’s contract
stipulates that the genotyping laboratories will report RFLP results
for 90% of isolates within 15 working days from the date of the
RFLP analysis request.
In almost all instances, these turnaround times are fast enough
to ensure that there will be no need for TB programs to prioritize
requests, even for investigations of outbreaks or false-positive
cultures.
Designation of Clusters and Reporting
Automated instruments determine the spoligotype and MIRU type.
The results (15-digit octal number for spoligotypes and 12-digit
number for MIRU types) are imported directly into the genotyping
laboratory-tracking database. The automated nature of the analyses
and assignment of numeric results reduces human interpretation and
transcription errors.
The spoligotype and MIRU type will be compared with all other isolates
previously submitted from the same TB program. The genotyping laboratory
will assign a PCR cluster designation to isolates that have spoligotypes
and MIRU types that match exactly. The PCR cluster designation will
consist of a two-letter TB program designation followed by a consecutively
derived three-digit number. For state programs, the two-letter designation
will be the postal code (e.g., “GA” for Georgia’s state TB program).
See Chapter 4, Combining Genotyping and Epidemiologic Data to
Improve Our Understanding of Tuberculosis Transmission, for
examples.
For each new isolate from a specific TB program, the search for
matches will yield one of three results:
- No match. This indicates that a search of current and
previous isolates from the TB program identified no other isolate
with a matching PCR genotype. Because the search for matches will
be restricted to isolates from the TB program, it is possible
that a matching isolate from another TB program exists in the
genotyping laboratory’s database. See Chapter 5, Developing
a Tuberculosis Genotyping Program, for a discussion of how
TB programs can compare genotyping results with other TB programs.
- A match with one current or previous isolate. The genotyping
laboratory will assign a new genotyping cluster number to this
newly identified cluster.
- A match with more than one isolate from an existing
genotyping cluster. The genotyping laboratory will assign the
existing cluster number to this newly identified addition to an
existing cluster.
When this matching analysis is complete, the results will be sent
by e-mail or other means to the TB program. At a minimum, the report
will list for each isolate the original isolate number supplied
by the submitting laboratory, the accession number assigned by the
genotyping laboratory, the report date, the spoligotype and MIRU
type, and cluster designation (for clustered isolates). For new
clusters, the report will list the previous isolate that matches
the new isolate so that the TB program can update their database.
These reports will be generated automatically, and each program
can specify any other data from the original submission form to
be included in the report. It is the responsibility of the TB program
to distribute the genotyping results as they see fit to others in
their jurisdiction.
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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