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Journal of Clinical Microbiology, August 2000, p. 3133-3134, Vol. 38, No. 8
0095-1137/00/$04.00+0
LETTERS TO THE EDITOR
The MB/BacT Is a Sensitive Method of Isolating
Mycobacterium tuberculosis from Clinical Specimens in a
Laboratory with a Low Rate of Isolation
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LETTER |
Roggenkamp and colleagues recently reported the results of
their comparative evaluation of the MB/BacT and the BACTEC 460 (3). They observed a significantly lower recovery rate for Mycobacterium tuberculosis using the MB/BacT, and they
commented that new diagnostic systems should be evaluated under
conditions found in nonspecialized laboratories that have a low
rate of detection of specimens positive for M. tuberculosis.
Our comparative study of these two systems (1) was performed
in a nonspecialized hospital laboratory that processes approximately 5,000 specimens per year for mycobacterial culture, with 5.6% positive
for mycobacteria and 1.5% positive for M. tuberculosis, similar to the numbers reported by Roggenkamp et al. We demonstrated that the MB/BacT detected the M. tuberculosis-M. avium
complex at a rate that was not significantly different from that of the BACTEC 460. The numbers of specimens that were culture positive for
mycobacteria in our study (44) were less than the 123 specimens positive in the study of Roggenkamp et al., and it is possible that with more specimens, differences may have been found. The decreased sensitivity of the MB/BacT system observed by Roggenkamp et
al. could be due to differences in the specimens studied, the products
used, and the methods. The epidemiology and presentation of patients
with mycobacterial disease in Europe and the United States could differ
and thereby influence the number of organisms in specimens. Our
specimens were predominantly of respiratory origin, whereas the
specimens in the Roggenkamp study included urine and gastric fluid, as
well as samples from sterile sites. Among 23 specimens positive for
M. tuberculosis in our study, 15 (65%) were smear
positive, in comparison to 25 of 71 (35%) in the Roggenkamp study.
Unlike Roggenkamp and colleagues, we screened all specimens with the
more sensitive Auramine O fluorochrome as mandated by the College of
American Pathologists (2). The MB/BacT detected all eight
smear-negative M. tuberculosis-infected patients in our
study. In a previous evaluation of the MB/BacT system, we had observed
decreased sensitivity for nontuberculous mycobacteria and a
high rate of nonmycobacterial contamination. The manufacturer
subsequently revised the medium and the antibiotic reconstitution
supplement. Our published study was performed with the revised
reconstitution fluid and antibiotic supplement in which oleic acid was
substituted for Tween 80 to enhance mycobacterial growth, and
vancomycin was added to reduce overgrowth of gram-positive bacteria.
Roggenkamp and colleagues used the original supplement. We maximized
the recovery of mycobacteria in the MB/BacT by staining contaminated
bottles weekly for the full 6 weeks of incubation. Twelve of 33 (36%)
of the mycobacteria not detected in the Roggenkamp study, 6 of
which were M. tuberculosis, were due to bacterial contamination that may have been improved by using the revised antibiotic supplement that is now available. The performance of this
revised supplement will depend on the handling of specimens and the
patient population studied.
In summary, the poor results obtained with the MB/BacT by Roggenkamp et
al. might have been influenced by the type of specimens studied,
how they were processed, and the growth medium and
antibiotic supplement utilized. It would be interesting for them to do
a study using the same type of experimental design and media as we
reported in our trial of this instrument.
 |
REFERENCES |
| 1.
|
Benjamin, W. H.,
K. B. Waites,
A. Beverly,
L. Gibbs,
M. Waller,
S. Nix,
S. A. Moser, and M. Willert.
1998.
Comparison of the MB/BacT system with a revised antibiotic supplement kit to the BACTEC 460 system for detection of mycobacteria in clinical specimens.
J. Clin. Microbiol.
36:3234-3238[Abstract/Free Full Text].
|
| 2.
|
Metchock, B. G.,
F. S. Nolte, and Richard J Wallace, Jr.
1999.
Mycobacterium, p. 399-437.
In
P. R. Murray, E. J. Baron, M. A. Pfaller, F. C. Tenover, and R. H. Yolken (ed.), Manual of clinical microbiology, 7th ed. American Society for Microbiology, Washington, D.C.
|
| 3.
|
Roggenkamp, A.,
M. W. Hornef,
A. Masch,
B. Aigner,
I. B. Autenrieth, and J. Heesemann.
1999.
Comparison of MB/BacT and BACTEC 460 TB systems for recovery of mycobacteria in a routine diagnostic laboratory.
J. Clin. Microbiol.
37:3711-3712[Abstract/Free Full Text].
|
| | | | |
William H. Benjamin Jr.
Ken B. Waites
Stephen A. Moser
Department of Pathology University of Alabama at Birmingham Birmingham, Alabama 35233-7331
|
 |
AUTHOR'S REPLY |
One topic
two studies
conflicting results? We think not! The
designs of the two studies have to be considered when considering the different results. The study of Benjamin and colleagues was not
designed to detect differences in sensitivity between the MB/BacT and
the BACTEC 460 when used in routine laboratories.
As stated by the authors themselves, their comparative study was
performed in a nonspecialized hospital laboratory that processes approximately 5,000 specimens per year for mycobacterial culture with
1.5% positive for M. tuberculosis. In their study, they
included a selection of 488 specimens, which were collected over a
4-month period and 5% of which were culture positive for M. tuberculosis (1). Likewise, the high percentage of
smear-positive specimens (65%) and the small number of specimens
collected per patient (1.61) indicated that their study material
was not representative for a routine laboratory. A study which excludes
certain clinical specimens from analysis cannot determine the
sensitivity of the assay in routine laboratories. As shown in both
studies, the cultivation of mycobacteria from smear-positive specimens
is not problematic when the MB/BacT and BACTEC 460 systems are used
(1, 2). The detection of M. tuberculosis in
smear-negative specimens (low quantity of mycobacteria) is important in
analyzing the sensitivity of the named systems. Benjamin and colleagues
presented only eight smear-negative specimens which were
culture-positive for M. tuberculosis. We included 46 in our study (2).
There is no doubt that the composition of the culture medium is
highly important in the diagnostic of mycobacteria. Supplementation with vancomycin reduces contamination. However, vancomycin itself has
antimycobacterial activity. Questions concerning the sensitivity of new culture media have to be answered in comparative studies in
which one detection system is used and identical specimens are
inoculated into different media that are to be compared.
Provided that the staining procedure is performed in the correct way,
the Ziehl-Neelsen stain is as sensitive as Auramine O fluorochrome in
detecting mycobacteria, although Ziehl-Neelsen staining is more
time-consuming. Our coworkers are well trained, not only in microscopy
and conventional cultivation procedures for mycobacteria but also in
the operation of the MB/BacT system (course of instruction and
periodical supervision by manufacturer coworkers).
 |
REFERENCES |
| 1.
|
Benjamin, W. H., Jr.,
K. B. Waites,
A. Beverly,
L. Gibbs,
M. Waller,
S. Nix,
S. A. Moser, and M. Willert.
1998.
Comparison of the MB/BacT system with a revised antibiotic supplement kit to the BACTEC 460 system for detection of mycobacteria in clinical specimens.
J. Clin. Microbiol.
36:3234-3238.
|
| 2.
|
Roggenkamp, A.,
M. W. Hornef,
A. Masch,
B. Aigner,
I. B. Autenrieth, and J. Heesemann.
1999.
Comparison of MB/BacT and BACTEC 460 TB systems for recovery of mycobacteria in a routine diagnostic laboratory.
J. Clin. Microbiol.
37:3711-3712.
|
| | | | |
Andreas Roggenkamp
Max von Pettenkofer-Institute for Hygiene and Medical
Microbiologie Ludwig Maximilians University Munich Pettenkoferstrasse 9a 80366 Munich, Germany
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Journal of Clinical Microbiology, August 2000, p. 3133-3134, Vol. 38, No. 8
0095-1137/00/$04.00+0