MTB Test Research Articles

Enhancing the Accuracy of Peripheral Pulmonary Tuberculosis Diagnosis: A Comparative Evaluation of CapitalBioTM Mycobacterium Nucleic Acid Detection Test and Mycobacterium tuberculosis Isothermal RNA Amplification Assay Using Endobronchial Ultrasonography with Guide Sheath.

The diagnosis of tuberculosis located in the peripheral zone remains challenging and requires ultrasound bronchoscopy-guided maneuvers. We assessed the precision of CapitalBioTM Mycobacterium nucleic acid detection test (CapitalBio MTB test) and Mycobacterium tuberculosis isothermal RNA amplification test (MTB-RNA) using endobronchial ultrasonography with a guide sheath (EBUS-GS) for peripheral pulmonary tuberculosis (PTB) and compared it with those of acid-fast bacilli (AFB) smear and MTB culture tests. This retrospective analysis included 287 patients suspected of peripheral pulmonary tuberculosis who underwent EBUS-GS examinations, medical examination results of AFB smears, MTB culture, CapitalBio MTB test, and MTB-RNA were analyzed. We evaluated the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and area under the receiver operating characteristic curve (AUC), and its diagnostic accuracy for peripheral PTB was evaluated in comparison with the final clinical diagnosis. The overall sensitivity, specificity, PPV, NPV, and AUC of CapitalBio MTB test were 44.83%, 100.00%, 100.00%, 54.07%, and 0.72, respectively; those of MTB-RNA were 22.99%, 100.00%, 100.00%, 45.75%, and 0.61, respectively, and those for parallel test (CapitalBio MTB test or MTB-RNA) were 46.55%, 100.00%, 100.00%, 54.85%, and 0.73, respectively. These values for AFB smear were 9.2%, 97.35%, 84.21%, 41.04%, and 0.53, respectively, and those of MTB culture were 31.03%, 100.00%, 100.00%, 48.50%, and 0.69, respectively. The CapitalBio MTB test showed the best diagnostic performance compared with AFB smear, MTB culture, and MTB-RNA assays and was similar to the parallel test (CapitalBio MTB test or MTB-RNA). The CapitalBio MTB test combined with EBUS-GS had satisfactory diagnostic accuracy for diagnosing peripheral PTB.

Assessing the utility of Truenat in extrapulmonary tuberculosis diagnosis – A NRL’s experience

BackgroundDiagnosis of extra pulmonary TB (EPTB) remains a big challenge. While data on utility of Xpert testing in EPTB diagnosis is enormous, there is limited data on Truenat MTB testing. AimIn this study we aimed to evaluate the usefulness of Truenat in EPTB diagnosis. Materials and methodsThe study included patients suspected and/or treated for EPTB located from Chennai district during the year 2021–2022. All processed EPTB samples were subjected to smear microscopy, culture and Truenat MTB testing. ResultsOf the 195 samples tested, 38 (19.4%) samples were positive for EPTB by any one of the diagnostic methods (smear, culture, microscopy). Out of these 38, 16 (42.1 %) were positive for MTB by Truenat and negative by Culture, 12 (31.5%) were positive by culture but negative by Truenat and 8 (21%) were positive by both Truenat and/or smear and culture. The sensitivity and specificity of the test was calculated with the composite reference standard (Culture (exclusion of colonies as positives), clinical conditions, and smear) and was found to be 60% and 100% respectively. ConclusionTruenat MTB test is a cost-effective rapid molecular test that can be used only for the diagnosis of presumptive EPTB and not on follow-up samples.

Estimating Limit of Detection of M. Tuberculosis DNA as an Analyte Released from a Chemically Coated Solid Matrix and Using a WHO-approved CB-NAAT Platform

The limit of detection of Mtb (Mycobacterium tuberculosis) bacilli was evaluated by detection of released DNA from a chemically treated cellulose matrix. Loopful of the Mtb cells were suspended in phosphate buffer (pH 6.8), DNA was extracted from it and quantified using a commercial Mtb real time PCR quantitation kit. Synthetic sputum, prepared as the matrix of choice was mixed with defined number of Mtb bacilli and spotted onto the cellulose matrix. Mtb DNA from the matrix was released into a DNA-release buffer and detected using a WHO-approved CB-NAAT detection platform. One hundred and fifty, 500 and 1000 but not 10 Mtb bacilli/mL of synthetic sputum could be detected by this protocol. The result underlined the potential of cellulose matrix as a potent, room temperature, Mtb-infected sputum transportation, storage and DNA-release device for sensitive detection of Mtb using Mtb Xpert Ultra CB-NAAT as the WHO-approved Mtb test of choice.

Accuracy of GenoQuick MTB test in detection of Mycobacterium tuberculosis in sputum from TB presumptive patients in Uganda.

Objective:The objective of the study was to determine the diagnostic performance of the GenoQuick MTB test on heated sputum against the conventional Lowenstein–Jensen Mycobacterium tuberculosis culture as the reference method for tuberculosis diagnosis.Introduction:Fast, reliable, and easy-to-use tests for tuberculosis diagnosis are essential to achieving the Sustainable Development Goal of diagnosing and treating 90% of tuberculosis patients by 2030. We evaluated the diagnostic performance of the GenoQuick MTB, a polymerase chain reaction–lateral flow test, in Uganda, a resource-constrained, high tuberculosis- and HIV-burden setting.Methods:Fresh sputum samples from presumptive tuberculosis patients at Mulago Hospital were tested for M. tuberculosis using smear microscopy, GenoQuick MTB test, and Lowenstein–Jensen culture. For the GenoQuick MTB test, mycobacterial DNA was extracted by heating sputum at 95°C for 30 min while DNA amplification and detection were done following the manufacturer’s protocol (Hain Lifescience, Nehren, Germany). Sensitivity, specificity, and kappa agreements were calculated against Lowenstein–Jensen M. tuberculosis culture as a reference test using STATA V12.Results:Of the 86 tested samples, 30.2% had culture-confirmed pulmonary tuberculosis. Overall, sensitivity was higher for GenoQuick MTB (81%, 95% confidence interval: 60%−93%) than for smear microscopy (69%, 95% confidence interval: 48%−86%). Among people living with HIV, sensitivity was identical for GenoQuick MTB and smear tests (75%, 95% confidence interval: 42%−95%). Contrastingly, smear had a higher overall specificity (98%, 95% confidence interval: 91%−100%) than for GenoQuick MTB (92%, 95% confidence interval: 81%−97%). A similar trend of specificity was observed among the people living with HIV for smear microscopy (100%, 95% CI: 87%−100%) and for GenoQuick MTB (96%, 95% confidence interval: 81%−100%).Conclusion:The GenoQuick MTB test could be a potential tuberculosis diagnostic test given its higher sensitivity. Evaluation of this test in larger studies is recommended.

Age and sex distribution of Mycobacterium tuberculosis infection and rifampicin resistance in Myanmar as detected by Xpert MTB/RIF

BackgroundDetection of tuberculosis disease (TB) and timely identification of Mycobacterium tuberculosis (Mtb) strains that are resistant to treatment are key to halting tuberculosis transmission, improving treatment outcomes, and reducing mortality.MethodsWe used 332,657 Xpert MTB/RIF assay results, captured as part of the Myanmar Data Utilization Project, to characterize Mtb test positivity and rifampicin resistance by both age and sex, and to evaluate risk factors associated with rifampicin resistance.ResultsOverall, 70% of individuals diagnosed with TB were males. Test positivity was higher among males (47%) compared to females (39%). The highest positivity by age occurred among individuals aged 16–20, with test positivity for females (65%) higher than for males (57%). Although a greater absolute number of males were rifampicin resistant, a greater proportion of females (11.4%) were rifampicin resistant as compared to males (9.3%). In the multivariate model, history of previous treatment, age less than 30, testing in the Yangon region, and female sex were significantly positively associated with rifampicin resistance after adjusting for HIV status and year test was performed.ConclusionsOur results indicate that young adults were more likely to test positive for TB and be identified as rifampicin resistant compared to older adults.

Lipoarabinomannan Antigen Assay (TB-LAM) for Diagnosing Pulmonary Tuberculosis in Children with Severe Acute Malnutrition in Mozambique.

BackgroundTuberculosis (TB) and malnutrition are important causes of morbidity and mortality in children in the developing world.AimsTo assess the prevalence of pulmonary TB in severely malnourished children and evaluate TB detection using the urine lipoarabinomannan antigen assay (TB-LAM).MethodsA retrospective analysis was conducted in all pediatric inpatients with severe acute malnutrition at a rural health center in Mozambique, from February to August 2018. All children underwent a physical examination and chest X-ray, and their nasopharyngeal aspirates and stool specimens were studied for mycobacterial culture and subjected to the Xpert MTB/RIF assay. TB-LAM tests were performed on urine.ResultsOf 45 included cases, 17 (37.8%) were clinically diagnosed as pulmonary TB. None of these were detected by the Xpert MTB test; 4 (8.9%) nasopharyngeal aspirates were TB-culture positive. Seventeen patients (37.8%)—all clinically diagnosed with TB—tested positive on the TB-LAM, while 23 (51.1%) were negative. In 5 (11.1%), the urine LAM was not done.ConclusionAlthough our sample size was small, TB was diagnosed and treated in more than a third of included children. The urine TB-LAM test showed a perfect correlation with clinical diagnosis of childhood TB.LAY SUMMARYSevere acute malnutrition makes children more vulnerable to tuberculosis (TB) infections, but it is difficult to detect TB in children because they cannot always cough up phlegm, which is used in diagnostic processes. This study aimed to find out how many severely malnourished children had TB in Gaza, Mozambique, and to test the accuracy of a less-used diagnostic test: the lipoarabinomannan assay (TB-LAM). Of the 45 severely malnourished children who were admitted to our hospital, 17 were diagnosed with TB by their doctor. The TB-LAM corroborated the clinical diagnosis in all cases, while the other tests (Xpert MTB/RIF assay) and cultures failed to detect most of them. Overall, more than a third of severely malnourished children had TB, and the TB-LAM test—a simple, point-of-care method—was a highly accurate way to diagnose them. While larger studies are needed to confirm these results, our findings suggest that the TB-LAM could vastly improve TB diagnosis in malnourished children.

Neurotuberculosis in an immunocompetent patient

Neurotuberculosis is a serious disease, occurring in about 5 to 10% of cases of extrapulmonary tuberculosis. The clinical spectrum varies according to the affected site (meningeal, cerebral parenchyma or spinal cord). Herein we present the case of a 32-year-old male patient with cough and weight loss who presented with hemiparesis and disorientation, receiving a diagnosis of neurotuberculosis using the Xpert MTB / RIF® test. The present case aims to report the clinical and neuroimaging data of an immunocompetent patient with neurotuberculosis.The definitive diagnosis is made by the detection of tuberculosis bacilli in cerebrospinal fluid. The RIF® Xpert MTB test has similar sensitivity and specificity to other diagnostic methods for tuberculosis and the advantage of providing information on resistance to rifampicin. The prognosis depends on the stage and onset of treatment.

Adhesion to zirconia as a function of primers/silane coupling agents, luting cement types, aging and test methods

This study evaluated the adhesion of resin cements to zirconia with different primers/silane coupling agents using two test methods with and without aging. Zirconia discs (Cercon) (N = 900, n = 15 per group) were ground finished to 2000 grit silicone carbide and randomly divided into seven groups: (a) C: No treatment (Control), (b) SG: Signum, (c) CL: Clearfil Ceramic Primer, (d) AP: Alloy Primer, (e) Monobond Plus, (f) ES-R: ESPE-Sil after Rocatec and (g) ES-C: ESPE-Sil after CoJet. Methacrylate (Variolink II-VL) and MDP based (Panavia F2.0-PN) dual-polymerized and self-adhesive resin cements (RelyX Unicem-RX) were adhered and polymerized accordingly. The specimens were further randomly divided into two groups to be tested after (a) 24-h dry storage at 37 °C and (b) thermocycling (×5000, 5–55 °C). Macroshear (MSB) and macrotensile bond tests (MTB) were conducted in an universal testing machine (crosshead speed: 1 mm/min) and failure types were analyzed after debonding. Data were analyzed using Univariate analysis and Tukey’s tests (α = 0.05). Two-parameter Weibull modulus, scale (m) and shape (0) were calculated. While primer/silane (p < 0.001), cement type (p < 0.001) and aging (p < 0.001) significantly affected the bond results, test method did not show significant difference (p = 0.237). In MSB test, Weilbul moduli were more favorable for MP-VL (4.2) and AP-PN (6) combinations and after aging for MP-VL (4.2) and AP-PN (5.66). In MTB test, after aging, Weilbul moduli were more favorable for AP-PN (5.41). Bond strength results mostly decreased with SG (24–92%) after aging. Cohesive failures in the cement were more frequent with PN (252) compared to VL (83).

Early detection of Mycobacterium tuberculosis complex in BACTEC MGIT cultures using nucleic acid amplification.

We evaluated the application of nucleic acid amplification (NAA) in liquid cultures for the early detection of Mycobacterium tuberculosis. The Cobas TaqMan MTB test, IS6110 real-time PCR, and hsp65 PCR-restriction fragment length polymorphism (RFLP) analysis were used to detect BACTEC MGIT 960 (MGIT) cultures on days 3, 5, 7, and 14. The procedure was initially tested with a reference strain, H37Rv (ATCC 27294). Subsequently, 200 clinical specimens, including 150 Acid Fast bacillus (AFB) smear-positive and 50 AFB smear-negative samples, were examined. The Cobas TaqMan MTB test and IS6110-based PCR analysis were able to detect M. tuberculosis after 1day when the inoculum of H37Rv was >3 x 10(-2) CFU/ml. After a 5-day incubation in the MGIT system, all three NAA assays had a positive detection regardless of the inoculum size. After a 1-day incubation of the clinical specimens in the MGIT system, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for the Cobas TaqMan MTB assay were 70.2%, 100%, 100%, and 82.3% respectively. For IS6110-based PCR analysis, these values were 63.1%, 100%, 100%, and 78.9%, and were 88.1%, 100%, 100%, and 92.1% respectively for hsp65 PCR-RFLP analysis. After a 3-day incubation, the specificity and PPV were 100% for all three NAA tests; the Cobas TaqMan MTB assay had the best sensitivity (97.6%) and NPV (98.3%). The sensitivity, specificity, PPV, and NPV for conventional culture analysis were 98.8%, 100%, 100%, and 99.1%. Thus, NAA may be useful for the early detection of M. tuberculosis after 3 days in MGIT.

Evaluation of the Cobas TaqMan MTB test for detection of Mycobacterium tuberculosis complex

Background: The Cobas TaqMan MTB assay is used for rapid detection of the Mycobacterium tuberculosis complex (MTC) in clinical samples. It is only validated for respiratory samples, but is often requested by physicians for non-respiratory specimens. The aim of this study was therefore to evaluate the performance of this assay in clinical praxis in a country with low prevalence of tuberculosis (TB). Methods: The results from 2388 respiratory and 1005 non-respiratory human specimens were analyzed by this real-time PCR technique. Using culture results as the gold standard, the sensitivity, specificity, positive predictive value (PPV), and negative predictive values (NPV) of the PCR results were calculated. Results: For smear-positive respiratory specimens all the four values investigated were 100%. The number of smear-positive non-respiratory specimens was only eight, and all of these eight specimens reacted positively. Sensitivity was 51% for smear-negative respiratory specimens, and 47% for smear-negative non-respiratory specimens. When all non-respiratory specimens were analyzed together the sensitivity was 51%. Specimens from 16 patients were PCR-positive but culture-negative and these cases are discussed. In one case, TB DNA was still present in sputum 2 years after a successful treatment. Conclusion: The study shows that the Cobas TaqMan MTB assay performs well in smear-positive samples, while the sensitivities are unsatisfactory for both respiratory and non-respiratory smear-negative specimens. Furthermore, the analyses emphasize that this assay cannot be used to evaluate treatment or contagiousness, or to detect relapses or screen for TB.

Evaluation of a modified direct agar proportion method for testing susceptibility of Mycobacterium tuberculosis from MGIT samples

The emergence of resistance to anti-tuberculosis (TB) drugs has become an obstacle to effective TB control. Thus, there is an urgent need to identify patients and initiate adequate treatment for drug-resistant cases in a timely manner. The BACTEC MGIT 960 system is well known for its rapid culturing time, and is in widespread use in Taiwan. In this study, we evaluated the possibility of replacing the traditional indirect agar proportion method with a modified direct agar proportion method (MDAPM), as a technique for rapid testing the drug susceptibility of Mycobacterium tuberculosis without additional cost. In this study, 432 positive MGIT 960 samples that were identified as M. tuberculosis complex using the MeDiPro M. tuberculosis Antigen Rapid Test or the Cobas Amplicor MTB test were evaluated. Each sample was tested separately by the MDAPM and indirect agar proportion method, between July 2008 and December 2008, to compare the consistency and total turnaround time. Four first-line anti-TB drugs-rifampin, isoniazid, ethambutol, and streptomycin-were tested. For the MDAPM and indirect agar proportion method, the respective consistencies for each drug were 99.31%, 98.38%, 98.38%, and 97.22%. Our results also indicated that the MDAPM leads to an average saving in working time of 2 weeks, compared with the traditional indirect agar proportion method. In addition to having the potential to shorten turnaround time without compromising diagnostic quality, the MDAPM also provides a more efficient and cost-effective procedure. This modified procedure presents potential benefits for TB diagnosis in laboratories already equipped with the MGIT 960 system.

Comparison of the Xpert MTB/RIF and Cobas TaqMan MTB Assays for Detection of Mycobacterium tuberculosis in Respiratory Specimens

The Xpert MTB/RIF assay (Cepheid, Sunnyvale, CA) is a fully automated, cartridge-based real-time PCR assay designed to detect Mycobacterium tuberculosis and rifampin resistance within 2 h. The performance of the Xpert assay has been evaluated in various clinical settings. However, there are few data comparing the Xpert assay to the Cobas TaqMan MTB test (Roche Diagnostics, Basel, Switzerland), one of the most widely utilized molecular assays for M. tuberculosis detection. In this prospective study, 320 consecutive respiratory specimens were processed simultaneously using acid-fast bacillus (AFB) staining, mycobacterial cultures with both solid and liquid media, and the Cobas and Xpert assays. The Xpert assay was performed with direct respiratory specimens, while the Cobas assay was done with decontaminated concentrated specimens. Based on the culture as a reference method, the overall sensitivities of the Cobas and Xpert assays were 71.4% and 67.9%, respectively. When AFB smear results were taken into consideration, the sensitivities of the Cobas assay for smear-positive and -negative specimens were 87% and 54%, while those of the Xpert assay were 67% and 69%, respectively. The Cobas assay showed 100% specificity and 100% positive predictive value (PPV) regardless of smear results, while the Xpert assay showed 100% specificity and 100% PPV for smear-positive specimens but 98% specificity and 60% PPV for smear-negative specimens. In conclusion, the Xpert assay showed performance that was slightly inferior to that of the Cobas assay but seems useful for the rapid detection of M. tuberculosis, considering that it was performed without laborious and time-consuming decontamination and concentration procedures.

Clinical value of IS6110-based loop-mediated isothermal amplification for detection of Mycobacterium tuberculosis complex in respiratory specimens

A fundamental to global tuberculosis (TB) control is timely and accurate diagnosis of infectious cases of the disease. Among various methods, techniques based on nucleic acid amplification are the ones with promising prospects. The present study evaluates the diagnostic value of the recently developed IS6110-based loop-mediated isothermal amplification (LAMP) for detection of Mycobacterium tuberculosis complex (MTBC) in sputum specimens. In this cross-sectional study (2008-2009), IS6110-LAMP was evaluated on 101 sputum specimens from 93 highly suspected TB patients and compared to Amplicor MTB test and in-house IS6110-PCR and -nested PCR assays. Culture results or clinical recovery following anti-TB therapy was considered as a reference to prove the TB cases. The overall sensitivity of IS6110-LAMP, Amplicor, nPCR, and PCR were respectively 89.6% (69/77 specimens; 95% confidence interval [CI], 80.5-95.4%), 76.6% (59/77 specimens; CI, 65.6-85.5%), 79.2% (61/77 specimens; CI, 68.5-87.6%) and 59.7% (46/77 specimens; CI, 47.9-70.8%). The specificity and positive predictive value (PPV) were 100% for all the tests, and the negative predictive value (NPV) of IS6110-LAMP, Amplicor, nPCR, and PCR were respectively 75%, 57.1%, 60%, and 43.6%. There was an excellent overall agreement between LAMP and nPCR (k 0.828), and between LAMP and Amplicor (k 0.746), in addition to a better tolerance of IS6110-LAMP to inhibitors present in clinical specimens. The better diagnostic performance of IS6110-LAMP compared to Amplicor (p = 0.009), nPCR (p = 0.013) and PCR (p < 0.0001) besides its rapidity, simplicity, and cost-effectiveness makes it a valuable method for the detection of MTBC in clinical samples, particularly in resource-limited settings.

Evaluation of Real-time Polymerase Chain Reaction for Detection of the 16S Ribosomal RNA Gene of Mycobacterium tuberculosis and the Diagnosis of Cervical Tuberculous Lymphadenitis in a Country With a High Tuberculosis Incidence

Tuberculous lymphadenitis (TBL) is the most common form of extrapulmonary tuberculosis. Currently, the standard diagnostic test for TBL is culture, which takes more than several weeks to yield results. We studied a real-time polymerase chain reaction (PCR) for rapid detection of Mycobacterium tuberculosis in cervical lymph node specimens obtained from patients in a country where the tuberculosis incidence is high. Patients with cervical lymphadenopathy were prospectively enrolled between April 2009 and March 2010. Clinical specimens obtained through fine-needle aspiration (FNA) and excisional biopsy were tested for M. tuberculosis by the COBAS TaqMan MTB Test, a real-time PCR assay for detecting the 16S ribosomal RNA gene of M. tuberculosis. Mycobacterial culture and histopathological findings from tissue biopsy specimens were used as a reference standard for sensitivity and specificity calculations. Of 73 patients, 41 received a diagnosis of TBL. For biopsy specimens, the sensitivity of real-time PCR was 63.4%, and the specificity was 96.9%. For FNA specimens, the sensitivity was 17.1%, and the specificity was 100%. The sensitivity of real-time PCR of biopsy specimens was comparable to that of tissue culture but significant lower than that of histopathological examination (P < .01). Real-time PCR did not increase the yield for rapid diagnosis of TBL.

Removal of PCR Inhibitors in Real-time PCR for Mycobacterium tuberculosis

Background: The inhibition rates for nucleic acid tests of Mycobacterium tuberculosis have been reported to range from less than 1% to more than 10%. Specimen dilution, boiling, addition of bovine serum albumin (BSA), and a silica membrane can be used to override amplification inhibitors in nucleic acid tests of M. tuberculosis. The inhibition rate for real-time PCR of M. tuberculosis (COBAS TaqMan MTB test; Roche Diagnostics, Manheim, Germany) and effective strategies to override PCR inhibitors were investigated in this study. Methods: The inhibition rate for COBAS TaqMan MTB test was investigated in 980 clinical specimens. The effectiveness of PCR inhibitor removal by repeated run, dilution, boiling, addition of BSA, and use of silica membrane were evaluated in the inhibited specimens. Results: Inhibitory substances were present in 4.1% of specimens (40/980). Among 40 inhibited specimens, inhibitory substances were removed in 12 (30%), 30 (75%), 27 (67.5%), 25 (62.5%) and 12 (30%) specimens with repeated run, dilution, addition of RBS, boiling and use of silica membrane, respectively. Conclusion: The overall inhibition rate for the COBAS TaqMan MTB test was 4.1%. Dilution, boiling and addition of BSA were shown to be more effective than repeated run and use of silica membrane for removal of PCR inhibitors. A combination of two methods might be useful and should be studied in the future. (Korean J Clin Microbiol 2011;14:97-102)

Evaluation of the Cobas TaqMan MTB Test for Direct Detection of Mycobacterium tuberculosis Complex in Respiratory Specimens

The Cobas TaqMan MTB test, based on real-time PCR technology, was evaluated for direct detection of Mycobacterium tuberculosis complex (MTBC) in respiratory specimens. A total of 1,093 samples from 446 patients, including 118 acid-fast smear-positive and 975 acid-fast smear-negative specimens, were investigated. Diagnostic cultures performed with 7H11 agar, Löwenstein-Jensen medium, and the Bactec MGIT 960 system were considered the reference methods. When discrepant results between the Cobas TaqMan MTB test and culture occurred, additional results from the BD MGIT TBc identification test and the GenoType Mycobacterium CM test performed on growth-positive and acid-fast-stain-positive MGIT tubes and review of the patient's medical history were used for discrepancy analysis. The overall sensitivity, specificity, positive predictive value, and negative predictive value for the Cobas TaqMan MTB test were 91.5%, 98.7%, 91.5%, and 98.7%, respectively. In general, the performance of the new Cobas TaqMan MTB test was comparable to that of the replaced Cobas Amplicor MTB system. The most prominent feature of the new system was its extraordinarily high sensitivity (79.5%) for detecting MTBC in smear-negative specimens; out of 44 smear-negative but culture-positive specimens, 35 were positive by the new system. The Cobas TaqMan MTB assay, including DNA extraction, can be completed within 3 h.

Evaluation of Cobas TaqMan MTB PCR for Detection of Mycobacterium tuberculosis

Nucleic acid-based amplification tests allow the rapid detection of Mycobacterium tuberculosis. Recently, a real-time PCR assay for M. tuberculosis complex, the Cobas TaqMan MTB test (Roche Diagnostics, Basel, Switzerland), was introduced. We performed a prospective study to evaluate the diagnostic performance of the Cobas TaqMan MTB test system. A total of 406 specimens collected from 247 patients were simultaneously tested by conventional culture, Cobas Amplicor MTB PCR, and TaqMan MTB PCR. The cross-reactivity with other Mycobacterium species and the detection limit were also evaluated. Among 406 specimens, a total of 24 specimens (5.9%) were culture positive: 14 specimens were positive by both TaqMan and Amplicor MTB PCRs, while 5 specimens were positive by only TaqMan PCR. The remaining five specimens were negative by both PCR methods. Seven specimens with negative culture results were positive by TaqMan PCR, but five of these were negative by Amplicor MTB PCR. The sensitivity, specificity, and positive (PPV) and negative (NPV) predictive values were 79.1%, 98.2%, 73.1%, and 98.7% for TaqMan and 58.3%, 99.5%, 87.5%, and 97.4% for the Amplicor MTB PCR test, respectively. There was no cross-reactivity with M. tuberculosis and nontuberculous mycobacterial species. The detection limit for the Cobas TaqMan MTB PCR test was 4.0 copies/μl. The Cobas TaqMan MTB PCR test showed higher sensitivity for detection of the M. tuberculosis complex without disturbing the specificity and NPV than the Amplicor MTB PCR test.

Assessment of the Amplicor PCR for the detection of Mycobacterium tuberculosis in smear negative respiratory and non respiratory specimens: a retrospective analysis

Commercial polymerase chain reaction (PCR) kits are widely accepted for analysis of smear positive respiratory specimens, but the sensitivity is variable for smear negative ones. To assess the PCR method usefulness in smear negative respiratory and non respiratory specimens. We compared the PCR results (AMPLICOR MTB test, Roche) of 235 specimens subjected to culture in Loewenstein-Jensen agar (as the gold standard). 181 (76%) were respiratory and 54 (24%) extra-respiratory specimens. The sensitivity was 88%) and 50%>, respectively, specificity and PPV was 100%> in both cases. NPV was 99.4%> in respiratory specimens and 96.1% in non-respiratory specimens. The good performance of this PCR in smear negative respiratory specimens allows the clinician to take decisions based on the result of this exam. In extra-respiratory specimens the contribution is important only when the PCR result is positive.

Análisis retrospectivo del rendimiento de Amplicor-PCR® para la detección de Mycobacterium tuberculosis en muestras respiratorias y no respiratorias con baciloscopia negativa: A retrospective analysis

Commercial polymerase chain reaction (PCR) kits are widely accepted for analysis of smear positive respiratory specimens, but the sensitivity is variable for smear negative ones.To assess the PCR method usefulness in smear negative respiratory and non respiratory specimens.We compared the PCR results (AMPLICOR MTB test, Roche) of 235 specimens subjected to culture in Loewenstein-Jensen agar (as the gold standard).181 (76%) were respiratory and 54 (24%) extra-respiratory specimens. The sensitivity was 88%) and 50%>, respectively, specificity and PPV was 100%> in both cases. NPV was 99.4%> in respiratory specimens and 96.1% in non-respiratory specimens.The good performance of this PCR in smear negative respiratory specimens allows the clinician to take decisions based on the result of this exam. In extra-respiratory specimens the contribution is important only when the PCR result is positive.

Nontuberculous mycobacterial infections among patients suspected of pulmonary tuberculosis

The purpose of this study was to present a retrospective analysis of the frequency of nontuberculous mycobacteria (NTM)-related pulmonary infections among the AFB-positive and/or culture-positive patients in the Warsaw region who were suspected of tuberculosis (TB) and hospitalized in the university hospital between 1999 and 2005. All the AFB-positive pulmonary samples were examined with a molecular method using the Amplicor MTB test (Roche) for detection of Mycobacterium tuberculosis complex, and all mycobacterial isolates were speciated by high performance liquid chromatography (HPLC) analysis of mycolic acids. Patients who met clinical, radiological, and bacteriological criteria of mycobacteriosis were classified according to the American Thoracic Society (ATS) guidelines for diagnosis of NTM related disease. Among the 445 smear-positive or/and culture-positive patients, 142 subjects (31.9%) were found to be infected with M. tuberculosis. Among 303 non-TB patients, mycobacteriosis was found in 27 (8.9%) subjects. The frequency of NTM-related lung disease as compared to the bacteriologically-confirmed lung TB was estimated at 1:5. The rapid, precise methods of NTM speciation are necessary for progress in diagnostics of NTM related diseases.