In this study, PCR-RFLP analysis (PRA) of the gene encoding 65-kD heat shock protein (hsp65) was used for identification of 70 clinical NTM isolates obtained from two mycobacteriology laboratories in Malaysia. Approximately one third of the isolates were identified as M. fortuitum. A total of 24.3% and 10% of the isolates were identified as M. abscessus and M. avium complex respectively. Other isolates identified were M. kansasii, M. marinum, M. simiae, M. parascrofulaceum, M. szulgai, and M. gordonae. Six clinical isolates were untypable by this approach. Sequence analysis of the 439-bp region of the hsp65 gene and a 564-bp region of the 16S rRNA gene revealed high degrees (97%-100%) of sequence similarity of these isolates with ''M. insubricum'', M. terrae, M. senegalense/M. conceptionense and M. houstonense in the public sequence databases. This study reported the identification of mycobacteria species which are considered rare or new to this region. As NTM are found widely in the environment, the clinical significance of the isolates in this study is yet to be determined.
Background: Nocardiosis is a clinical and diagnostic challenge, compounded by lacunae in existing literature. Our objectives were to establish the clinical spectrum of this disease in our setting, describe the most common causative agent of the disease and to ascertain differences in our patient population from available data.Methods & Materials: This was a 10 year (2004)(2005)(2006)(2007)(2008)(2009)(2010)(2011)(2012)(2013) retrospective study carried out at a tertiary care centre in South India, of 131 cases of nocardiosis. The electronic medical records were studied and data analysed.Results: Sixty three percent were male, 23% of all in the sixth decade of life. The most common sites of infection were the skin and the eye -36 (27%) patients each and the lower respiratory tract -35 patients(26%). 48 (37%) patients were on immunosuppressant therapy, either a triple drug therapy following renal transplant, autoimmune disorders/ haematological malignancies on combination immunosuppressants or patients on prolonged corticosteroids. Of 36 patients with nocardiosis of the eye, 30 (83%) were corneal ulcers with history of trauma with vegetative matter or soil, and 5(14%) were endophthalmitis following intraocular lens implantation. 16(46%) patients with respiratory tract nocardiosis had a previous lung pathology. 11(8%) were HIV associated nocardiosis. Disseminated disease was seen in 7(5.3%) patients following renal transplant and in 3(2.3%) patients with SLE, all on triple drug immunosuppression. The most common organism isolated was Nocardia asteroides in 73(56%), followed by Nocardia spp in 32(24%), aerobic actinomycetes in 24(18%) and Nocardia brasiliensis in 2(1.5%). All patients responded to treatment with cotrimoxazole alone or in addition to surgical debridement for cutaneous and subcutaneous lesions. There was only one Nocardiosis related death in this cohort of patients. Antimicrobial susceptibility testing performed on 72 isolates showed 6.9% , 9.7%, 31%, 38%, 75%, 42%, 31%, 74% susceptibility to penicillin, ampicillin, erythromycin, tetracycline, cotrimoxazole, chloramphenicol, cefazolin and triple sulfa respectively. Conclusion:We report a predominance of nocardiosis from the eye and nocardiosis following immunosuppression. The most common species isolated was N.asteroides. A paucity in HIV associated nocardiosis is striking. Antimicrobial susceptibility showed 75% susceptibility to cotrimoxazole, the drug of choice, which was reflected by a good response to therapy in this cohort.
93.2% and 91.5%. The QFT had 58.3% sensitivity and 95.3% specificity in diagnosing children with latent TB. The commonest discordant results were TST + / QFT-in 15 of 141 children without TB, not unexpected in this BCG-vaccinated population. Conclusion:The QFT performed better than the TST in the diagnosis of tuberculosis. Although only moderately sensitive, they were highly specific in ruling out TB and showed good concordance in TB-negative children. Although a case may be made for using both tests in BCG-vaccinated children, the higher costs and technical expertise required for the QFT do not support its use instead of the cheaper and simpler TST in India.
Results: Blood cultures from 40 neonates received during the study period. Blood cultures from 18/40 neonates showed bacterial growth. Nine of 18 (50%) neonates showed the growth of E.faecium. All the E.faecium isolates were susceptible to Vancomycin and Linezolid. Susceptibility to other antibiotics :uniformly susceptible to Quinupristin/Dalfopristin and Chloramphenicol. Resistant to macrolides,fluoroquinolones,Gentamicin(high-level). Sequences of seven of the nine isolates were deposited in Gen-Bank (GenBank accession numbers HM222631 to HM222637). The sequence of the each isolate was different from the other. The neonates were either preterm or low birth weight. Babies presented with respiratory distress(6/9), with seizures (2/9) and refusal to feed (1/9).Conclusion: Among the neonates from whom Enterococcus faecium was isolated in blood, no specific clinical feature could be noticed. The isolates were found to be different from each other in our attempt to establish the relatedness of the strains.
e32413th International Congress on Infectious Diseases Abstracts, Poster Presentations Australia) detect interferon-gamma (IFN-␥) production in whole blood samples in response to stimulation with TB antigens. The role of QFT and Tuberculin skin test (TST) in the diagnosis of active TB among adults in high burden countries is not clear.Methods: We prospectively evaluated pulmonary and extrapulmonary TB suspects from a tertiary center in India, in a blinded comparison of new diagnostic tests. We aim to recruit 200 patients for the study. The blood samples collected from the patients were processed as per manufacturers instructions. The cut off for positivity used was 0.35 IU/ml. TST was performed using 2TU dose and 10 mm or greater was considered positive. Both were evaluated against a combined gold standard of solid (Lowenstein Jensen) and liquid (BACTEC 460 TB) culture.Results: To date the results of QFT and culture for 51 patients are available. Four indeterminate results were not included. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) of QFT for culture positive TB were 81% (54-95), 67% (47-82), 57% (35-76) and 87% (65-96) respectively. 52 patients had TST and culture results available. The sensitivity, specificity, PPV and NPV of TST for culture positive TB were 68% (45-85), 50% (32-68), 50% (32-68) and 68% (45-85) respectively.Conclusion: QFT has adequate sensitivity but poor specificity to detect active TB in India. QFT shows a trend to better sensitivity than TST. As expected, latent TB infection causes false positives. QFT is a single visit test with good negative predictive value but should not be used alone to rule out active TB.Background: Conventional cultures for TB have limitations. Solid media cultures are time consuming and liquid cultures are too expensive for developing countries. MODS is thought to be more sensitive, more rapid and less expensive than conventional culture because it detects early growth at a microscopic level.Methods: We prospectively evaluated pulmonary and extrapulmonary TB suspects who met inclusion criteria, from a tertiary center in India, in a blinded comparison of new diagnostics. We aim to recruit 200 patients for the study. Specimens were pretreated with NALC/NaOH and inoculated into 7H9 middlebrook liquid media with OADC and PANTA in a 24 well tissue culture plate and incubated at 37 • C incubator. The liquid media was observed daily using an inverted microscope for 'cording' growth typical of M. tuberculosis complex. MODS was evaluated against combined gold standard of solid and liquid (BACTEC 460 TB) culture.Results: The results of 67 patients are available to date and were included in this analysis. The sensitivity, speci-
Background: Tuberculosis (TB) is a communicable disease for which an early diagnosis is essential to control the disease. The microscopy-based TB screening is the conventional method employed for TB identification in sputum smears. Fluorescence microscopy-based diagnosis provides improved sensitivity and benefits large number of TB burdened communities across the globe. Microscopic images are often corrupted by intensity variations because of inherent imperfections of the image formation process. This may result in false positives which is the potential shortcoming of fluorescence microscopy.Methods & Materials: The fluorescence-stained slides were prepared at South African National Health Laboratory Services, Groote Schuur Hospital in Cape Town. The images (N=100) were captured using a camera in monochrome binning mode attached to a 20x objective fluorescence microscope of 0.5 numerical aperture. The camera (AxioCam HR) has a resolution of 4164 x 3120 with a pixel size 6.45 m (h) x 6.45 m (v).The illumination correction methods adopted in this work include surface fitting method, multiple regression method and bidirectional empirical mode decomposition. The results of illumination correction are validated using the image sharpness measures. This includes derivative-based, statistical, histogrambased and transform-based parameters.
Background: Rapidly growing mycobacteria (RGM) are environmental organisms that can cause post-operative wound infections. Infections typically occur after laparoscopic surgery due to inadequate sterilization of heat-sensitive instruments. We describe the clinical presentation and management of postoperative RGM infections at Christian Medical College (CMC), a large tertiary referral hospital in South India.Methods & Materials: Laboratory records from 1 st January 2012 to 31 st August 2015 were examined to identify patients with culture positive post-operative RGM infections. The electronic medical records of these patients were reviewed together with their haematological, histological and radiographic data.Results: Over this period, 32 patients were diagnosed with culture proven RGM infection as a consequence of surgery. Mycobacterium fortuitum was the commonest isolate (46.9%), followed by M. abscessus (31.2%) and M. chelonae (18.8%). Most patients had wound infections (96.9%); 78.1% extended into underlying muscle and 28.1% into structures deep to muscle. 37.5% patients had infection associated with prosthetic material including surgical mesh, pacemakers, cardiac valve and a neurosurgical shunt. Surprisingly, most patients (65.6%) had undergone open surgical rather than laparoscopic procedure (25%).Only 4 patients (12.5%) acquired RGM infection following surgery at CMC. Over this period, 96,713 operations were performed resulting in an infection rate of 0.004%. 87.5% patients underwent operation at a different hospital, presenting to CMC a median 4 months after operation. 43.8% received inappropriate treatment for wound infection before presenting to CMC. 37.5% received antibiotics and 9.4% empirical antitubercular therapy, highlighting poor knowledge about RGM infections.All patients were treated with surgical debridement; 75% received subsequent antibiotics consisting of a two or three drug combination of amikacin, levofloxacin/moxifloxacin, clarithromycin or linezolid. Patients jointly managed by surgeons and infectious disease physicians had a higher rate of clinical response (75%) with less loss to follow up (25%) than those managed exclusively by surgeons (43.8% and 57.25% respectively).Conclusion: RGM infections continue to complicate routine operations in India, although they are a rare complication of surgery in our hospital. They are under-recognised and frequently misdiagnosed resulting in delays in appropriate treatment. Higher clinical response rates are seen where management involves surgeons and infectious disease clinicians with laboratory support from microbiologists.
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