Market assessment of tuberculosis diagnostics in China in 2012.

OBJECTIVE: To assess the 2012 served available market for tuberculosis (TB) diagnostics in China in the sector served by the China Centre for Disease Control and Prevention (CDC) and the hospital sector in China, including both designated TB hospitals and general hospitals. DESIGN: Test volumes and unit costs were assessed for tuberculin skin tests, interferon-gamma release assays (IGRAs), smear microscopy, serology, cultures, speciation tests, nucleic-acid amplification tests (NAATs), drug susceptibility tests and adenosine-deaminase tests (ADA). Data were obtained from electronic databases (CDC sector) and through surveys (hospital sector), and were estimated for the two sectors and for the country as a whole. Test costs were estimated by staff at China CDC, and using published literature. RESULTS: In 2012, the China CDC and hospital sectors performed a total of 44 million TB diagnostic tests at an overall value of US$294 million. Tests used by the CDC sector were smear microscopy, solid and liquid culture and DST, while the hospital sector also used IGRAs, NAATs, ADA and serology. The hospital sector accounted for 76% of the overall test volume and 94% of the market value. CONCLUSION: China has a very large TB diagnostic market that encompasses a wide range of diagnostic tests, with the majority being performed in Chinese hospitals.

Market assessment of tuberculosis diagnostics in India in 2013.

BACKGROUND: India represents a significant potential market for new tests. We assessed India's market for tuberculosis (TB) diagnostics in 2013. METHODS: Test volumes and unit costs were assessed for tuberculin tests, interferon-gamma release assays, sputum smear microscopy, serology, culture, speciation testing, nucleic-acid amplification tests (i.e., in-house polymerase chain reaction, Xpert(®) MTB/RIF, line-probe assays) and drug susceptibility testing. Data from the public sector were collected from the Revised National TB Control Programme reports. Private sector data were collected through a survey of private laboratories and practitioners. Data were also collected from manufacturers. RESULTS: In 2013, India's public sector performed 19.2 million tests, with a market value of US$22.9 million. The private sector performed 13.6 million tests, with a market value of US$60.4 million when prices charged to the patient were applied. The overall market was US$70.8 million when unit costs from the ingredient approach were used for the 32.8 million TB tests performed in the entire country. Smear microscopy was the most common test performed, accounting for 25% of the overall market value. CONCLUSION: India's estimated market value for TB diagnostics in 2013 was US$70.8 million. These data should be of relevance to test developers, donors and implementers.

Routine use of the Gen-Probe MTD2 amplification test for detection of Mycobacterium tuberculosis in clinical specimens in a large public health mycobacteriology laboratory.

The new version of the Amplified Mycobacterium Tuberculosis Direct Test, MTD2 (Gen-Probe Inc., San Diego, CA) has been implemented as part of the regular testing algorithm for detecting Mycobacterium tuberculosis (MTB) in selected respiratory and non-respiratory specimens in our laboratory. At the Central Public Health Laboratory, Etobicoke, Ontario, we receive specimens for the detection of mycobacteria from all areas of the Province of Ontario. The laboratory processes approximately 25,000 specimens per year, and receives approximately 2000 reference cultures for identification. There are 600 to 700 new cases of tuberculosis detected yearly. Over the 1-year period (1997-98), 823 specimens were tested by MTD2 and the results were compared with radiometric culture (Bactec, Becton Dickinson, Sparks, MD) and clinical diagnosis, giving an overall sensitivity, specificity, positive predictive value and negative predictive values of 100%, 99.6%, 97.4% and 100%, respectively. Two hundred and two cases of respiratory TB and 56 cases of extrapulmonary TB were detected by MTD2 within 0-4 days of specimen arrival in the laboratory. By appropriate selection of specimens for testing, the MTD2 can provide a fast, accurate, and cost-effective method for the detection of MTB in clinical specimens.

Rapid molecular diagnosis of tuberculous meningitis using the Gen-probe Amplified Mycobacterium Tuberculosis direct test in a large Canadian public health laboratory.

OBJECTIVE: A 5-year retrospective study of the performance of the Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test (MTD) for detecting Mycobacterium tuberculosis complex in cerebrospinal fluid (CSF). Patient data from culture-confirmed cases of tuberculous meningitis (TBM) were also analysed. RESULTS: In total, 311 CSF specimens were tested by the MTD, of which 17 were positive. When compared with culture (gold standard), the sensitivity and specificity of the MTD test were 93.8% and 99.3%, respectively. The positive and negative predictive values for TBM were 88.2%, and 99.7%. Clinical and epidemiological information was requested for all culture-positive TBM patients. These data were used to assess the mortality rate (55.6%) and to determine common factors that could be applied as selection criteria for the appropriate testing of CSF by MTD. CONCLUSION: The study found the MTD test to be a rapid, sensitive and specific test for TBM. A history of immigration from an area endemic for tuberculosis (TB), a history of TB, symptoms of neurological deficits and the results of CSF analyses could be used to appropriately select CSF for MTD testing in order to provide a critical early diagnosis of TBM.

Investigation of tuberculosis transmission in Canadian Arctic Inuit communities using DNA fingerprinting.

OBJECTIVE: To understand the transmission of tuberculosis in Inuit communities in the Baffin region of the Canadian Arctic. METHODS: Twenty-one isolates of Mycobacterium tuberculosis from 19 Inuit patients diagnosed with tuberculosis between February 1991 and September 1993 were analyzed by DNA fingerprinting. The DNA fingerprints were achieved by the standard restriction fragment length polymorphism (RFLP) technique, with subsequent probing using the repetitive insertion segment IS6110. RESULTS: The isolates could be divided into three DNA types. The DNA types generally corresponded to the geographic origins of the patients. In most instances only one DNA type of M. tuberculosis was identified in each community. This suggests that a single case was the start of each of the three clusters, most likely due to reactivation. CONCLUSIONS: The results show that molecular typing of M. tuberculosis was useful in determining the mode of transmission of tuberculosis in a remote area of the Canadian Arctic where the disease is endemic. In addition, the information provides useful information for planning interventions in this setting.

Isolation of non-tuberculous mycobacteria among patients with pulmonary tuberculosis in Ontario, Canada.

SETTING AND OBJECTIVE: There are limited data regarding the frequency and significance of co-isolating pulmonary non-tuberculous mycobacteria (NTM) in patients with pulmonary tuberculosis (PTB). DESIGN: We identified all patients with culture-proven PTB in Ontario, Canada, in 2004, identified those with NTM 'co-isolation' (≤6 months following initial TB isolate) and determined subsequent NTM isolation over 5 years. RESULTS: In 2004, 369 people in Ontario had culture-proven PTB (average age 46 years, SD 21, 41% female). NTM co-isolation occurred in 11% (40/369), including Mycobacterium avium complex 22/40 (55%), M. xenopi 7/40 (18%), M. gordonae 6/40 (15%) and others 5/40 (13%). Patients with NTM co-isolation were older (55 vs. 45 years, P = 0.004), but had similar sex ratios (females 43% vs. 40%, P = 0.87). Among patients with co-isolation, 23% (9/40) went on to have ≥2 NTM cultures (excluding initial culture), compared with 3% (10/329) in the PTB group (including initial culture, P = 0.0001). In the co-isolation group, the median (quartiles) number of samples collected for mycobacterial study was 6 (4-8) compared to 2 (1-4) in the PTB group (P < 0.0001). CONCLUSIONS: The high frequency of subsequent NTM isolation among patients with NTM co-isolation during PTB may warrant follow-up for potential NTM disease.

Genotypic characterization of tuberculosis transmission within Toronto's under-housed population, 1997-2008.

Toronto has been the site of a recent extended tuberculosis (TB) outbreak in the homeless or under-housed population. Genotyping has identified a unique strain that continues to circulate within this population, with spread to individuals with no links to the shelter system, and anecdotally appears to progress rapidly from infection to active disease in some cases. The recent appearance and transmission of another unique strain was also identified, indicating that TB transmission continues to be a problem within the under-housed population. Enhanced surveillance utilizing molecular epidemiology is a useful tool to assist in TB control in vulnerable populations.

Use of gas chromatographic fatty acid and mycolic acid cleavage product determination to differentiate among Mycobacterium genavense, Mycobacterium fortuitum, Mycobacterium simiae, and Mycobacterium tuberculosis.

Three Mycobacterium genavense strains and three American Type Culture Collection reference strains each of Mycobacterium fortuitum, Mycobacterium simiae, and Mycobacterium tuberculosis were subcultured onto Mycobacteria 7H11 agar (Difco Laboratories, Detroit, Mich.) supplemented with mycobactin J (Allied Laboratories, Fayette, Mo.). After 4 weeks of incubation at 37 degrees C in 10% CO2, the cultures were analyzed by gas-liquid chromatography (GLC) for their fatty acids and mycolic acid cleavage products. M. fortuitum was clearly differentiated from M. genavense by the presence of the specific marker 2-methyloctadecenoic acid in M. fortuitum and by the ratio of tetracosanoic acid to hexacosanoic acid. This ratio was <1 for M. genavense and >3 for M. fortuitum. M. fortuitum also contained docosanoic acid, which was not detected in M. genavense. M. genavense, M. simiae, and M. tuberculosis, which have similar GLC profiles, were also differentiated from each other by the presence of either cis-10-hexadecenoic acid or cis-11-hexadecenoic acid and by tetradecanoic acid content.

Direct identification of Mycobacterium species in Bactec 7H12B medium by gas-liquid chromatography.

Twenty-nine Mycobacterium reference strains representing 10 species and 60 mycobacterial cultures isolated from sputum specimens were studied. These cultures were grown in Bactec 7H12B medium (Becton Dickinson and Co., Paramus, N.J.) supplemented with oleic acid-albumin-dextrose-catalase enrichment broth (Becton Dickinson and Co., Cockeysville, Md.). The cultures were analyzed by gas-liquid chromatography for their fatty acids, secondary alcohols, and mycolic acid cleavage products. All of the clinical isolates could be identified by comparing their gas-liquid chromatography profiles with those of the reference strains. The data indicate that this method significantly shortens the turnaround time and could be used for the early detection and identification of mycobacterial species.

Increasing drug resistance of Mycobacterium tuberculosis isolates in Ontario, Canada, 1987-1998.

We examined trends in resistance to first-line antituberculous agents for Mycobacterium tuberculosis strains isolated in Ontario, Canada from 1987 through 1998 (n=8069). The proportions resistant were as follows: isoniazid, 9.6%; rifampin, 1.9%; streptomycin, 4. 9%; ethambutol, 1.3%; and pyrazinamide, 1.7%. Resistance to isoniazid has increased markedly since 1990, whereas resistance to streptomycin, ethambutol, and pyrazinamide increased from 1997 through 1998. Resistance to both isoniazid and rifampin did not increase. The incidence of persistence and reactivation (early or late treatment failure) was 1-2 per 100 person-years (PY) in the first 7-12 months and 0.3-0.9 per 100 PY from 13 months to 5 years thereafter. For initially susceptible strains, the incidence of resistance to isoniazid was 0.11 per 100 PY and for and rifampin was 0.06 per 100 PY in the first year and negligible thereafter, with an overall risk of 0.14% for isoniazid and 0.10% for rifampin. Resistance of M. tuberculosis to antituberculous agents, and in particular to isoniazid, is a growing problem in Ontario and is higher than elsewhere in Canada.