BD MGIT" TBcIdentification Test

English: pages 1-5 Italiano: pagine 13- 16 8085917(03)

Français : pages 5- 8 Español: páginas 16- 20 2016-12
Deutsch: Seiten 9- 12

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pårstvi. I Naudojimo instrukcijs teiraukits vietos BD jgaliotojo atstovo. / Neem contact op met uw plaatselijke
BD-vertegenwoordiger voor instructies. / Kontakt din lokale BD-representant for mer informasjon. IAby uzyska
instrukcje u|ytkowania, skontaktuj si z lokalnym przedstawicielstwem BD. /Contacte o representante local
da BD para instruções. / Pentru instructiuni, contactaci reprezentantul local BD. / Ana nony4eHuA ykasaHMP
o6patuTecb KMecTHOMy npegcraBuTenio KOMnaHMnBD. l Inatrukcie získate umiestneho zástupcuspolonosti
BD. / Obratite se svom lokalnom predstavnikukompanije BD za uputstva. /Kontakta närmaste BD-representant
för anvisningar. /Talimatlar için yerel BD temsilcinizle temasa geçin. I3a iHCTpykuiAMM 3BepH0TbCA A0 MÍC;eBoro
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INTENDED USE
The BD MGITTM TBC ldentification Test (TBc ID) is a rapid chromatographic immunoassay for the qualitative detection
of Mycobacterium tuberculosis complex (MTbc) antigen from AFB smear-positive MGIT tubes. The device will detect the
following species of the MTbc: M. tuberculosis, M. bovis, M. africanum, and M. microti.
SUMMARY AND EXPLANATION
Tuberculosis is a chronic disease caused by an infection of M. tuberculosis. It is a major global public health problem and
one of the leading causes of death from infectious disease in the world.1 There were an estimated 9.2 million new cases
of tuberculosis in 2006 with 1.5 million deaths.2 Ninety-five percent of the estimated incidence occurs in developing
countries with 80 percent reported from 22 countries.3
Conventionalmethods for identification of mycobacteria rely on staining specimens for acid-fast bacili (AFB) followed
by culturing on solid media and biochemical testing. It can take as long as two months to speciate an isolate using these
standard methods.4 The World Health Organization (WHO) has recommended the use of liquid media for the culture of
M. tuberculosis.5 Modern procedures used to identify M. tuberculosis from liquid culture include nucleic acid probes and
gas-liquid /high performance chromatography.
It is important to distinguish non-tuberculous mycobacteria (NTM) from Mycobacterium tuberculosis complex (MTbc).
The BD MGIT TBC ID is a rapid procedure that identifies MTbc from culture grown in liquid media. It is a simple
chromatographic immunoassay which requires no sample preparation. The total assay time is 15 minutes with reactivity
determined by visual color development.
PRINCIPLES OF THE PROCEDURE
The TBc ID test isachromatographic immunoassay for the qualitative detection of MTbc from an AFB smear-positive
MGIT tube. This product detects MPT64, a mycobacterial protein fraction that is secreted from MTbc cells during culture.
When samples are added to the test device, MPT64 antigen binds to anti-MPT64 antibodies conjugated to visualizing
particles on the test strip. The antigen-conjugate complex migrates across the test strip to the reaction area and is
captured by asecond specific MPT64 antibody applied to the membrane. If the MPT64 antigen is present in the sample,
acolor reaction is produced by the labeled colloidal gold particles and is visualized as a pink to red line.
REAGENTS
Materials Provided:
The following components are included:
BD MGITTM TBC 25 devices Foil pouched device (containing a single reactive strip)and desiccant. The strip
ID test devices contains a test line of MPT64-specific monoclonal antibody and a control line of
anti-species antibody.
Materials Required But Not Provided: Pipettor (capable of delivering 100 pL), sterile pipette tips and timer.
Warnings and Precautions
For in vitro Diagnostic Use
1 Pathogenic microorganisms, including hepatitis viruses, Human mmunodeficiency Virus and novel influenza viruses,
may be present in clinical specimens. "Standard Precautions"%-9 and institutional guidelines should be followed in
handling, storing and disposing of all specimens and allitems contaminated with blood and other body fluids.
2 Do not use devices beyond the expiration date.
3 Do not reuse the device.
 4 Use a clean, sterile pipette tip for each sample.
body fluid, tissue, sputa and bronchial lavage fluid).
5. Do not test clinical specimens directly in this device (e.g., human
Procedure should be carried out using
6 Use appropriate biosafety precautions for handling mycobacteria.
appropriate biological safety cabinets.4.8
Used devices should be discarded according
7. Used test devices may contain viable MTbc which could be infectious.
to your institutional guidelines or Standard Precautions requirements.
MGIT system for the growth
Processed sputum and other non-sterile specimens are typically cultured in the
primary cultures can contain non-AFB microorganisms. If non-AFB
and detection of mycobacteria. As a result, metabolism may interfere with the BD MGIT
organisms predominate in a positive MGIT culture their growth and
with polymicrobic cultures containing both AFBand
TBC ID performance. Care must be taken when using this test AFB-positive organisms
non-AFB organisms.As a guideline, only use the TBc ID test on a positive MGIT tube if containingan
predominate on the smear. Performing the test on a very turbid positive culture (>1.0McFarland)
overgrowth of non-AFB organisms may cause a false positive result.
detectable limits. 10
9 There are reports of rare strains of MTbc that produce MPT64 antigen below
10. False positive results may be observed in the presence of protein A-producing strains of bacteria
(e.g. Staphylococcus aureus).
Danger

H228 Flammable solid.

P280 Wear protective gloves/protective clothing/leye protection/face protection. P370+P378 In case of fire: Use for
extinction: CO2, powder or water spray.
Storage and Handling: Test devices may be stored at 2-35 °C. DO NOT FREEZE. Devices must be at ambient room
temperature at time of testing.
SAMPLE COLLECTION AND PREPARATION
This test is designed to identify MTbc from AFB smear-positive MGIT tubes (4 mL and 7 mL). The presence of AFB in a
positive MGIT tube should be confirmed using an AFB smear prior to conducting the test.
Sample Storage: Positive MGIT tubes can be stored at 2-37 °C for up to 10 daysafter MGIT tube positivity and prior to
testing with the TBc ID device. If necessary. positive MGIT tubes may be stored and maintained at -20 to 8C for up to
two months.

PROCEDURE
NOTES:
AFB smear-positive MGIT tubes can be tested in the TBc ID device within 10 days after MGIT tube positivity.
If devices are refrigerated, they must be brought to ambient room temperature in the foil pouch prior to testing.
1. Remove the TBc ID device from its foil pouch immediately before testing. Place the device on a flat surface.
2 Label one device for each sample to be tested.
3 Thoroughly mix the sample (AFB smear-positive MGIT tube) by inverting or vortexing. Do not centrifuge.
4 Remove cap from MGIT tube and using a sterile pipette tip, pipette 100uL of the sample into the sample well
(as indicated by the teardrop ) of the appropriately labeled device. Tightly replace cap on MGIT tube. Start timer
for 15 min.
5. Read result at 15 min and record test result. Do not interpret test after 60 min.
QUALITY CONTROL
Each device contains both positive and negative internallprocedural controls. The appearance of a control line in the
read window at the Control "C" position provides an internalpositive control that validates the proper reagent function
and assures that the correct test procedure was followed. The membrane area surrounding the test and control lines
is the internal negative control for the device. Abackground area that is white to light pink indicates that the test is
performing correctly.
Additional quality control should be performed in accordance with local or country regulations, laboratory accreditation
requirements,and your laboratory's standard quality control procedures.
Positive and negative external controls should be tested in the same manner as test samples to provide a means of
externalquality control. Positive Control: Apositive MGIT tube prepared by growing a known isolate of MTbc. This should
yield a positive result. Negative Control: An uninoculated MGIT tube. This should yield a negative result.
External controls should be run, at a minimum, for each new lot or each new shipment received.
If the controls do not perform as expected, do not report sample results. Contact your local BD representative or
Technical Services for assistance.
usinglavage
fluid).
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JRETATION OF RESULTS
Positive Test for TBc (MPT64 antigen present) -A pink to red line appears at the lestT
position and the Control "C" position in the read window. This indicates MPT64 antigen was
detected in the sample. The intensity of the C and Tlines may vary. The background areashould
be white to light pink.

Negative Test for TBc (no MPT64 antigen detected) - No pink to red line is visible at the Test "T
read window. This indicates that MPT64 antigen was not detected in the sample. A
position of the
proper performance of the test procedure.
line at the Control "C" position read window indicates
The background area should be white to light pink.

is visible at the Control "C" position in the read

Invalid Test -The test is invalid if no pink to red line If invalid, the sample must be
interpretation.
window or if the background area color inhibits test
retested with a new device.

Invalid
REPORTING OF RESULTS
Positive Test Should be reported as MTb complex (MTbc).
Negative Test Should be reported as Acid-Fast Bacilli, non-MTbc.
Invalid Test Do not report results.
LIMITATIONS OF THE PROCEDURE
mycobacterial or mixed bacterial infections.
1 This test does not rule out the presence of other
organisms.
2 This test is unable to differentiate between MTb complex (MTbc)
of MTbc infection. The test results are to be used in
3 This test should not be used solely for the determination evaluation and other diagnostic procedures.
conjunction with information available from the patient's clinical
detect
possibility of infection with MTbc. The device is unable to
4 A negative result does not always rule out the results are to be used in conjunction with information
MTbc when a mutation arises in the MPT64 gene. The test
procedures.
available from the patient's clinical evaluation and other diagnostic
produce no MPT64 antigen and will therefore
5 Some substrains of M. bovis BCG among M. tuberculosis complex
result in anegative test result with the device.10

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PERFORMANCE CHARACTERISTICS
Cinical Studies
The BD MGIT TBc ID test was compared to a molecular identification method (Gen-Probe AccuProbe M MTh
ldentification Test) at one clinical site in a TB high burden country using a total of 247 AFB smear-positive, BD MCGIT
instrunment postive samples. AccuProbe identified 121 as MTbc and 126 as NTM. Acomparison of these results is shcoun
below in Table 1.
Table 1: Performance Summary of the BD MGIT TBc ID Test compared to AccuProbe with Percent Agreerments
and Confidence Intervals (CI)
AccuProbe

BD MGIT TBc ID
120 9
117

(+) =Positive (-)= Negative

"Six of the nine positive by TBc ID and negative by AccuProbe were positive by the
Hain GenoTypeTM MTBDR Plus.
Positive Percent Agreement (95% Cl): 99% (95.5%, 100%)
Negative Percent Agreement (95% CI): 93% (86.9%, 96.7%)
Overall Percent Agreement (95% CI):96% (92.7%, 98%)
Reproducibility
Reproducibility testing was performed at the clinical site using 6 negative samples and 6 positive sanples (containing
recombinant MPT64 antigen) for a total of 108 replicates. The overallreproducibility for the BD MGIT TBc ID test was 100%.
Analytical Studies
Analytical Specificity (Cross Reactivity): 756 samples of 23 non-MTb complex mycobacteria species were tested with no
cross reactivity. Fifty-three samples of 10 non-mycobacterial species (common respiratory contaminants) were tested: 3
of the non-mycobacterial samples caused interference at very high levels of MGIT tube contamination (>1.0 McFariand).
Internal Studies
The BD MGIT TBc ID test was evaluated using mycobacterial strains that were seeded into BD MGIT 960 (7 mL) and
Manual MGIT (4 mL) tubes. A total of 828 samples consisting of 23 MTbc strains (414 devices) and 23 NTM strains (414
devices) were tested. One device produced an invalid result.
Table 2: Performance Summary of the BD MGIT TBC ID Test Evaluated with MTbcand NTM Strains with
Percent Agreements
Mycobacteria Strains
BD MGIT TBc ID MTbc NTM
413

414
(+) = Positive (-)= Negative
Positive Percent Agreement: 100% (413/413)
Negative Percent Agreement: 100% (414 |414)
Overall Percent Agreement: 100% (827 | 827)

AVAILABILITY

Cat. No. Description

245159 BDMGIT TM TBC ldentification Test 25 Test Devices
245111 BD BBL TM MGIT TM 4 mL tube 25 tubes
245113 BD BBL TM MGIT TM 4 mL tube 100 tubes
245122 BDBBL TM MGIT IM 7 mL tube 100 tubes
REFERENCES
1 Centers for Disease Control and Prevention. 2008. Morbidity and Mortality Weekly Report Vol 57, No 11
2 World Health Organization. 2008. WHO Report 2008, Global Tuberculosis Control, Surveillance, Planning.
Financing. Geneva.
3 World Health Organization. 2007. WHO Report 2007, Global Tuberculosis Control, Surveillance, Planning.
Financing. Geneva.
4 Kent, P.T., and G.P. Kubica. 1985. Public health mycobacteriology: a guide for the level ll laboratory. U. S.
Department of Public Health and Human Services, Atlanta, GA.
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