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Siddiqi S.,Becton Dickinson | Ahmed A.,Indus Hospital | Asif S.,Indus Hospital | Behera D.,LRS Institute of TB and Respiratory Diseases | And 9 more authors.
Journal of Clinical Microbiology | Year: 2012

Conventional indirect drug susceptibility testing of Mycobacterium tuberculosis with liquid medium is well established and offers time-saving and reliable results. This multicenter study was carried out to evaluate if drug susceptibility testing (DST) can be successfully carried out directly from processed smear-positive specimens (direct DST) and if this approach could offer substantial time savings. Sputum specimens were digested, decontaminated, and concentrated by the laboratory routine procedure and were inoculated in Bactec MGIT 960 as well as Lowenstein-Jensen (LJ) medium for primary isolation. All the processed specimens which were acid-fast bacterium (AFB) smear positive were used for setting up direct DST for isoniazid (INH) and rifampin (RIF). After the antimicrobial mixture of polymyxin B, amphotericin B, nalidixic acid, trimethoprim, and azlocillin (PANTA) was added, the tubes were entered in the MGIT 960 instrument using the 21-day protocol (Bactec 960 pyrazinamide [PZA] protocol). Results obtained by direct DST were compared with those obtained by indirect DST to establish accuracy and time savings by this approach. Of a total of 360 AFB smear-positive sputum specimens set up for direct DST at four sites in three different countries, 307 (85%) specimens yielded reportable results. Average reporting time for direct DST was 11 days (range, 10 to 12 days). The average time savings by direct DST compared to indirect DST, which included time to isolate a culture and perform DST, was 8 days (range, 6 to 9 days). When results of direct DST were compared with those of indirect DST, there was 95.1% concordance with INH and 96.1% with rifampin. These findings indicate that direct DST with the Bactec MGIT 960 system offers further time savings and is a quick method to reliably detect multidrug resistance (MDR) cases. Copyright © 2012, American Society for Microbiology. All Rights Reserved.

Myneedu V.P.,LRS Institute of TB and Respiratory Diseases | Sharma P.P.,LRSI Institute of TB and Respiratory Diseases
Indian Journal of Tuberculosis | Year: 2011

Introduction: A large number of tuberculosis cases are continuously being reported from India and other developing countries leading to high morbidity and mortality. In spite of many newer tests available for diagnosing a case of tuberculosis, smear microscopy of sputum is still the preferred test under programmatic conditions. The current national and international guidelines recommend two sputum smear examinations in two days for diagnosing cases of tuberculosis, which is time-consuming, tedious, needs multiple visits, leading to high dropout of infectious cases. In the background of existing limitations of smear microscopy, we attempted to complete the diagnosis of tuberculosis on same day by serial collection of the spot sputum specimen and analyze its advantages, feasibility and viability. Material & Methods: The study was undertaken by the Department of Microbiology, Lala Ram Sarup Institute of Tuberculosis and Respiratory Diseases during May 2010 to April 2011. Sputum specimens were collected from 330 randomly selected tuberculosis suspects who attended OPD of hospital, patients submitted spot and home collected morning sputum sample in a standard method and spot and additional spot sputum(X- spot) collected one hour after the first spot sample as per the proposed front loading method. All the samples received were stained by acid fast Ziehl-Neelsen (ZN) stain and examined on the same day. The sputum sample was pooled and cultured in Lowenstein Jensen (U) media in duplicate set of bottles. The results of two different microscopic methods were compared with the gold standard culture test. Results: Out of the total 330 TB suspects, 70.60% were males and 29.39% females. The most common complaint was of cough with sputum (88.18%), chest pain (70.21%), fever (55.15%) and loss of appetite (43.03%). Upon examining the total sputum slides, 18.48 % were positive for acid fast bacilli. The smear positivity was 61/330(18.48%) by standard methods and in proposed new method 43/330(13.03%). Sensitivity of the standard and proposed new method smear microscopy was 58.25% and 40.07% respectively and specificity was 99.55% in both the methods. Conclusion: Same day smear microscopy for diagnosing tuberculosis by a proposed new method of smear examination in the case of suspected tuberculosis seems not a promising step towards improving the quality of sputum smear examination. The results of sensitivity and specificity of the two approaches were not similar. More than eighty per cent responded in favour of same day sputum delivery system and getting result on same day. This study can be confirmed on larger scale and preference of patients can be examined in peripheral laboratory also before taking it up for consideration in the national tuberculosis programme.

Visalakshi P.,LRS Institute of TB and Respiratory Diseases | Meharwal S.K.,Saba University | Myneedu V.P.,LRS Institute of TB and Respiratory Diseases | Behera D.,LRS Institute of TB and Respiratory Diseases
Diagnostic Microbiology and Infectious Disease | Year: 2010

The aim of this study was to evaluate a simple, rapid, and inexpensive colorimetric nitrate reductase assay (NRA) for direct drug susceptibility testing (DST) of Mycobacterium tuberculosis against rifampicin (RIF) and isoniazid (INH). A total of 118 smear-positive specimens were processed from patients on antituberculosis treatment. A comparison was made between the direct NRA of DST with the direct proportion method and with the internationally accepted indirect 1% proportion method as the "gold standard". The sensitivity and specificity of the direct NRA and indirect proportion method were 94% and 98%, and 100% and 98% for RIF and INH, respectively. Excellent agreement was found between the 2 tests with κ values of 0.92 and 0.98, and P value was less than 0.001 for RIF and INH. The results in most cases were available in 14 days (turnaround time). The direct NRA is a rapid, accurate, simple, and inexpensive method to determine multidrug resistance from sputum. Direct NRA may become an appropriate alternative method, especially for the resource poor settings. © 2010 Elsevier Inc. All rights reserved.

Kalra M.,Jawaharlal Institute of Postgraduate Medical Education & Research | Khuller G.K.,Jawaharlal Institute of Postgraduate Medical Education & Research | Grover A.,Jawaharlal Institute of Postgraduate Medical Education & Research | Behera D.,Lrs Institute Of Tb And Respiratory Diseases | And 2 more authors.
Diagnostic Microbiology and Infectious Disease | Year: 2010

We evaluated the diagnostic potential of a cocktail of 4 antigens encoded by regions of difference (RD) 1 and 2 of Mycobacterium tuberculosis, that is, early secretory antigenic target-6, culture filtrate protein-10 (CFP-10), CFP-21, and mycobacterial protein from species tuberculosis-64 (MPT-64) on the basis of antigen and antibody detection by enzyme-linked immunosorbent assay. Parallel detection of antigens and antibodies in the serum samples of pulmonary tuberculosis (PTB) patients resulted in higher sensitivity as compared to either of the single tests in both smear-positive (90%) and smear-negative (60%) PTB patients. In addition, combined detection of antigens and antibodies in the fluids of extrapulmonary tuberculosis (EPTB) patients could detect >90% of the patients with high specificity. These results demonstrate the ability of the combination of antigen and antibody detection assays based on the cocktail of RD antigens to diagnose a substantial number of PTB and EPTB cases with high specificity. © 2010 Elsevier Inc. All rights reserved.

Mehta P.K.,Maharshi Dayanand University | Kalra M.,Jawaharlal Institute of Postgraduate Medical Education & Research | Khuller G.K.,Jawaharlal Institute of Postgraduate Medical Education & Research | Behera D.,Lrs Institute Of Tb And Respiratory Diseases | Verma I.,Jawaharlal Institute of Postgraduate Medical Education & Research
Diagnostic Microbiology and Infectious Disease | Year: 2012

Immuno-polymerase chain reaction (I-PCR) combines the versatility of enzyme-linked immunosorbent assay (ELISA) with the exponential amplification power of PCR. The present study was designed to detect antibodies to Mycobacterium tuberculosis complex-specific region of difference (RD) antigens, i.e., early secretory antigenic target-6, culture filtrate protein-10, culture filtrate protein-21, and mycobacterial protein from species tuberculosis-64, as well as antigens in pulmonary tuberculosis patients by I-PCR assay. We could detect ESAT-6 and other RD antigens up to 0.1 fg by I-PCR assay, thus resulting in 10 7 times higher sensitivity than that observed with ELISA. With paired sample analysis based on the detection of antibodies in serum and antigens in sputum of the same individual, the sensitivity of RD multi-antigen cocktail-based I-PCR assay was 72% in smear-negative cases and 91% in smear-positive cases of pulmonary tuberculosis with high specificity values. In extrapulmonary tuberculosis patients, higher sensitivity was observed by detecting cocktail of antigens by I-PCR assay as compared to sensitivity earlier observed in the same samples by ELISA. © 2012 Elsevier Inc..

Srivastava V.,Immunology Group | Vashishta M.,LRS Institute of TB and Respiratory Diseases | Gupta S.,Immunology Group | Singla R.,LRS Institute of TB and Respiratory Diseases | And 4 more authors.
Immunology and Cell Biology | Year: 2011

Protective immune responses during Mycobacterium tuberculosis (M. tuberculosis) infection are regulated at multiple levels and critically dependent on the balance in the secretion of pro-inflammatory and regulatory cytokines. A key factor that governs this balance at the cellular level is suppressors of cytokine signaling (SOCS). We recently demonstrated that toll-like receptor 2 and dendritic cell (DC)-SIGNR1 differentially regulate SOCS1 expression in DCs during M. tuberculosis infection. This consecutively regulated IL-12 production and determined M. tuberculosis survival. In this study, we characterized the role of SOCS1 in regulating effector responses from CD4 + and CD8 + T cells during M. tuberculosis infection. Our data indicate that T cells from M. tuberculosis-infected mice show increased and differential association of SOCS1 with CD3 and CD28, when compared with uninfected mice. While SOCS1 displays increased association with CD3 than CD28 in CD4 + T cells; SOCS1 is associated more with CD28 than CD3 in CD8 + T cells. Further, SOCS1 shows increased association with IL-12 and IL-2 receptors in both CD4 + and CD8 + T cells from infected mice when compared with naive mice. Silencing SOCS1 in T cells increased signal transduction from T cell receptor (TCR) and CD28 with enhanced activation of key signaling molecules and proliferation. Significantly, SOCS1-silenced T cells mediated enhanced clearance of M. tuberculosis inside macrophages. Finally, adoptive transfer of SOCS1-silenced T cells in M. tuberculosis-infected mice mediated significant reduction in M. tuberculosis loads in spleen. These results exemplify the negative role played by SOCS1 during T cell priming and effector functions during M. tuberculosis infection. © 2011 Australasian Society for Immunology Inc. All rights reserved.

Sharma P.,LRS Institute of TB and Respiratory Diseases | Kumar A.,MD University | Singh P.,National Statistical Commission
Indian Journal of Community Medicine | Year: 2010

Background: Worldwide, the case notification rate of tuberculosis has been reported to be higher for men than women. In India also, the prevalence of TB is higher among males as compared to females but it is important to study the trend of gender gap in the prevalence of tuberculosis over the years. Objective: To examine the trend in gender gap in the prevalence of TB over the years. Materials and Methods: The unit level data of NFHS-2 (1998-99) and NFHS-3 (2005-06) has been utilized. Gender gap in the prevalence of TB has been estimated for the two rounds of the surveys. The delta (D), the difference in gender gap in two surveys, has been estimated and decomposed by background characteristics such as place of residence(urban/rural), religion (Hindus/Muslims/others), caste(SC/ST/OBC/others) and standard of living(SLI) (low/medium/high) categories. Main Findings: Overall, the prevalence of TB has remained almost same in the two surveys [432/lakh in NFHS-2 and 418/lakh in NFHJS-3; Z=1.19, P0 =0.275. The gender gap has increased to 217/lakh in NFHS-3 in comparison to 145 per lakh in NFHS-2. The increase in gender gap is significantly higher in rural areas [of 98 per lakh;167/ lakh in NFHS-2 vs 265/lakh in NFHS-3; P<0.05] as compared to corresponding increase in urban areas [of 30 per lakh; 88/ lakh in NFHS-2 vs118/ lakh in NFHS-3, P>0.05]. The increase in delta (D) (difference in gender gap in two surveys) is accounted for as 88% by the rural areas and 12% by the urban areas. Conclusion: The increase in gender gap in the prevalence of TB is more in rural areas as compared to urban areas. The increase in rural areas is mainly contributed by Hindus, SC and ST and low and medium SLI categories and in urban areas, the contribution is mainly by Hindus, other castes and high SLI categories.

Gupta K.,LRS Institute of TB and Respiratory Diseases | Thakur S.,LRS Institute of TB and Respiratory Diseases
Indian Journal of Pathology and Microbiology | Year: 2011

Malakoplakia of lung is an unusual condition that has been reported to occur in association with immunocompromised state, particularly in those with acquired immunodeficiency syndrome. We present two cases of pulmonary malakoplakia in immunocompetent individuals. The diagnosis was made on histopathological examination of surgically resected specimen.

Mathur M.K.,LRS Institute of TB and Respiratory Diseases | Verma A.K.,LRS Institute of TB and Respiratory Diseases | Makwana G.E.,P.A. College | Sinha M.,P.A. College
Journal of Global Infectious Diseases | Year: 2013

Introduction: Intestinal parasites predominantly coccidian parasites are a common cause for diarrhea in human immunodeficiency virus (HIV)-positive patients. Materials and Methods: The study was conducted during January 2009-December 2010. A total of 1,088 stool samples from 544 seropositive HIV positive cases were examined microscopically for ova and cyst using wet mount preparations and stained smears. Out of 544 patients, 343 had prolonged diarrhea for more than 4 weeks, 57 had acute diarrhea of lesser than 7 days and 144 were asymptomatic cases who attended out-patient department; included in this study after taking consent from patients. Enteric pathogens were detected in 274 (50.36%) of the 544 patients. Results and Conclusions: The parasites identified were Cryptosporidium (135), Isospora belli (42), Cyclospora (12), Microsporidia (02), Entamoeba histolytica (49), Hookworm (34). Intestinal parasites in chronic diarrhea were significantly higher than the acute diarrhea (63.05% vs. 7.35%; P < 0.05). Parasitic pathogens were frequently associated with HIV-positive patients with diarrhea in Western India. Stools of all HIV-positive patients with diarrhea should thoroughly be investigated to identify etiologic agents for proper management.

Dewan R.K.,LRS Institute of TB and Respiratory Diseases
European Journal of Cardio-thoracic Surgery | Year: 2010

Background: Surgery for pulmonary tuberculosis (TB) has become rather limited. However, it is still required for some sequelae and complications. This is a 15-year retrospective study of cases operated upon for pulmonary TB at a centre. Patients and methods: A total of 2878 cases underwent surgical procedures for various complications of pulmonary TB over a 15-year-long period. After excluding those managed by tube thoracostomy, rib resection and open-window thoracostomy, 1297 cases out of this series were taken up for major thoracic surgical procedures. A total of 98 were operated for persistent sputum-positive status, 740 for recurrent massive haemoptysis or chest infections, 2024 for empyema and 18 for diagnostic reasons. Procedures were 830 lung resections, 12 primary thoracoplasties, 295 space-reducing thoracoplasties, 158 decortications, 744 open-window thoracoplasties and 837 tube thoracotomies alone. Results: There were 18 early deaths and 37 late deaths. The cause of death was haemorrhage in seven cases and respiratory failure in nine cases and septicaemia in two cases. Late deaths were mostly because of progressive tubercular disease. There was significant morbidity in terms of broncho-pleural fistula (BPF) in 95 cases and persistent sinus in 37 cases. Milder complications such as pneumonia, fever and wound sepsis were noticed in some cases but definite records were not available. BPF was managed by tube drainage followed by either window thoracostomy or thoracoplasty. In multi-drug-resistant (MDR) cases, persistent documented sputum negativity was achieved in 64 out of 86 cases. Results were better in haemoptysis and chest infection group where the desired result was achieved in 699 cases. Conclusions: Surgery in pulmonary TB is still relevant in many cases and yields a very gratifying result. It is a challenging surgery and this series is a very large one. © 2009 European Association for Cardio-Thoracic Surgery.

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