Diagnostic Potential of Circulating Biomarkers in Adenosine Deaminase Diagnosed Pleural Tuberculosis Cases.

Pleural tuberculosis accounts for nearly 20% of Extra pulmonary tuberculosis. Adenosine deaminase, commonly used biomarker for the diagnosis, is non specific and there is paucity of literature on its correlation with conventional or newer methods for the diagnosis of extra pulmonary forms of TB. The aim of the study was to assess diagnostic potential of T cell function markers [interferon (IFN-γ), interleukin (IL-2) and IFN-γ/IL-2 ratio]; macrophage activation marker [neopterin]; and oxidative stress markers [protein carbonyl and malondialdehyde (MDA)] in pleural tuberculosis. 26 pleural TB cases diagnosed on the basis of suggestive chest X-ray and raised serum ADA levels and healthy controls were included in the study. Pleural fluid specimens were subjected to Zeihl Neelsen staining and culture on Lowenstein Jensen medium. Serum IFN-γ, IL-2, neopterin and protein carbonyl levels detection were done by ELISA and MDA levels were determined by measuring the thiobarbituric acid reactive substances. Median serum levels of IFN-γ, IL-2, IFN-γ/IL-2 ratio, neopterin, protein carbonyl and MDA were significantly different between cases and controls. Levels of all biomarkers except IL-2 were significantly higher in cases with contact history. Mean levels of ADA and ESR were 46.27 U/L and 46.62 mm/hr in PTB cases. AUC for IFN-γ, IL-2, IFN-γ/IL-2 ratio, neopterin, protein carbonyl and MDA were significantly discriminative for cases and controls. IFN-γ/IL-2 ratio was best discriminatory biomarker with highest area under ROC curve. Though no correlation was seen between ADA and any of the six biomarkers, ESR levels correlated significantly with all biomarkers except IL-2 by spearman's correlation coefficient. Though all the circulating biomarkers under study provide useful supportive evidence for the diagnosis of PTB, further studies involving diverse control groups particularly non-PTB effusion are needed to validate these results.

Neopterin and oxidative stress markers in the diagnosis of extrapulmonary tuberculosis.

BACKGROUND: Extrapulmonary tuberculosis (EPTB) often presents with nonspecific signs and symptoms. Further the paucibacillary nature of extrapulmonary specimens and irregular distribution of bacilli lower the sensitivity of conventional diagnostic methods making EPTB, a diagnostic dilemma. OBJECTIVE: To study neopterin, protein carbonyl and malondialdehyde (MDA) in EPTB. METHODS: Sixty nine clinically confirmed cases with an equal number of age and sex matched healthy controls were enrolled. Ziehl-Neelsen staining for acid fast bacilli and culture on Lowenstein-Jensen medium were performed on all the extrapulmonary specimens. Serum neopterin and protein carbonyl levels were estimated using commercial ELISA kits. Malondialdehyde was determined by measuring thiobarbituric acid reactive substances. RESULTS: Serum neopterin, protein carbonyl and MDA levels were significantly discriminative for cases of EPTB from healthy controls (p < 0.05). Levels of all the three biomarkers under study significantly differed between culture as well as smear positive and negative cases. A positive correlation between neopterin and protein carbonyl was seen among the cases. CONCLUSIONS: So far few studies have integrated combination of validated host biomarkers for active disease in EPTB. Our study suggests the potential diagnostic role of neopterin, protein carbonyl and MDA in EPTB.

Comparative Diagnostic Utility of Neopterin and IFN-γ/IL-2 in Extrapulmonary Tuberculosis.

Resurgence of TB has emphasized the need for newer methods of diagnosis. Extrapulmonary tuberculosis (EPTB), being paucibacillary, is a diagnostic dilemma. The aim of the present study was to correlate IFN-γ/IL-2 with neopterin in diagnosis of EPTB. Extrapulmonary specimens from 69 clinically diagnosed cases were stained by Ziehl-Neelsen and cultured on Lowenstein-Jensen medium for Mycobacterium tuberculosis. ELISA was used to assess serum IFN-γ, IL-2 and neopterin levels. Median serum levels of IFN-γ/IL-2 and neopterin were 3.22 and 21.6 nmol/L in clinically diagnosed EPTB cases and 0.52 and 4.20 nmol/L in healthy controls respectively (p < 0.001). Both IFN-γ/IL-2 and neopterin were significantly higher in culture positive (14.64 and 49.8 nmol/L) than culture negative cases (3.01 and 17.5 nmol/L) respectively (p < 0.05). IFN-γ/IL-2 was significantly higher in AFB smear positive cases (8.63) than smear negative cases (3.04) (p = 0.003), whereas no significant difference in neopterin levels was seen (p = 0.307). A positive correlation between IFN-γ/IL-2 and neopterin was seen in EPTB cases (spearman's rho = 0.453, p < 0.001), whereas in healthy controls no such correlation existed (spearman's rho = 0.018, p = 0.884). An urgent need for research in the field of biomarkers exists to utilize them as point of care test in the diagnosis of EPTB.

Correlation between the clinico radiological heterogeneity and the immune-inflammatory profiles in pediatric patients with neurocysticercosis from a tertiary referral centre.

INTRODUCTION: Neurocysticercosis (NCC) is the leading cause of epilepsy in developing world. Cysticercal lesions develop in brain depending upon a combination of host immune-inflammatory response, mainly mediated by cytokines produced by cysticercal antigens. AIM AND OBJECTIVES: To correlate between MRI findings and levels of Th1/Th2 cytokines present in sera of children clinically suspected of NCC with generalized or partial seizure. MATERIAL AND METHODS: Fifty children presenting with history of seizures and/or mass effects and/or hydrocephalous, with a diagnosis of NCC based on the clinical and radiological profile were included. Antibody (IgM) for NCC and Th1/Th2 cytokine response (TNF-α, IL-2/and IL-6) detection was done on sera from all the patients following manufacturer's instructions. RESULTS: Out of 50 cases, 10 presented with acute symptoms of NCC with an immunological response of a predominance of pro-inflammatory cytokines (IL-2: 8, TNF-α: 2). High IL-6 was found in 40 children indicating an active lesion with chronic granulomas suggestive of parasitic destruction and persisting presentation with seizures. However, the levels of IL-6 differed with values lower in patients with inactive (calcified lesions) forms of NCC. A significant proportion (43 of 50 cases) had negative serology, probably because of the waning of antibody response months or years after the parasites die. CONCLUSION: Parasite maintains equilibrium with host immune response in early infection, a mild Th1 response is provoked; but later this equilibrium is disturbed toward Th2 response that leads to parasite destruction. Number or stage of the parasites along with immunegenetic aspects may explain the pleomorphic and unpredictable course of NCC.

T helper 1 to T helper 2 shift in cytokine expression: an autoregulatory process in superantigen-associated psoriasis progression?

Psoriasis is an inflammatory skin disorder characterized by increased activation of CD4(+) T lymphocytes, and systemic and local overexpression of pro-inflammatory cytokines such as interleukin 2 (IL-2), gamma interferon (IFN-gamma), IL-6 and tumour necrosis factor alpha, indicating that immunopathogenesis of the disease is T helper 1 (Th1) mediated. Several studies suggest a pivotal role of bacterial superantigens in the initiation and/or exacerbation of this illness. This study was conducted to assess the systemic Th1/Th2 imbalance in Indian psoriasis patients presenting with variable duration of disease by studying systemic superantigen-stimulated peripheral blood mononuclear cell (PBMC) cytokine expression. PBMCs were isolated and stimulated in vitro with superantigens (streptococcal pyrogenic exotoxin A and staphylococcal enterotoxin B), and the cytokines released (IFN-gamma for a Th1 response, and IL-4 and IL-10 for a Th2 response) were assayed. In contrast to controls, psoriasis patients in the early course of disease were characterized by significantly increased expression of the pro-inflammatory cytokine IFN-gamma, whilst a shift towards IL-10 secretion (Th2 response) was observed in those presenting with increased duration of disease. These observations suggest a possible shift from a Th1 to a Th2 cytokine response with superantigen-associated progression for the duration of psoriasis, perhaps as an adaptive process by the immune system in an attempt to downregulate abnormal inflammatory Th1 immune responses.

Trend of antibiotic resistance of Vibrio cholerae strains from East Delhi.

BACKGROUND & OBJECTIVE: Epidemics of cholera caused by toxigenic Vibrio cholerae O1 and O139 (Bengal strain) represent a major public health problem in most developing countries. In view of the reported shift in epidemiology and pattern of antibiotic resistance in this was study carried out to assess the development of resistance to essential drugs like fluoroquinolones during treatment of cholera and cholera like cases in Delhi. METHODS: Faecal specimens collected from 1184 patients with cholera and cholera like illness between 2001-2006 admitted to Guru Teg Bahadur hospital, East Delhi were subjected to culture isolation. Antimicrobial susceptibility testing of V. cholerae isolates was done by disc diffusion method. RESULTS: Of the 1184 faecal samples examined, 670 (56.6%) were positive for V. cholera from 2001- 2006. V. cholerae El Tor Ogawa (54.6%) was more common than serotype Inaba (32.5%). During 2004-2006 V. cholerae Inaba emerged as the predominant serotype. Resistance to nalidixic acid, furazolidone and co-trimoxazole was constantly high (100%). Multiple antibiotic resistance (MAR) V. cholerae O1 Inaba isolates exhibited increased resistance to ciprofloxacin with MIC >4 microg/ml, but largely all remained susceptible to other antibiotics like, gentamicin, tetracycline and chloramphenicol. INTERPRETATION & CONCLUSION: V. cholerae have a permanent existence in the environment and during the quiescent period, their survival in water bodies allows dissipation of resistance patterns to different serotypes or strains of V. cholerae O1 and therefore there is need for constant observation.

Anti-HBs Titers Following Pentavalent Immunization (DTwP-HBV-Hib) in Term Normal Weight vs Low Birthweight Infants.

OBJECTIVE: To compare anti-HBs titers between term low birth weight (1800-2499 g) infants and normal birthweight infants, 6 weeks after last dose of primary immunization with pentavalent vaccine, and to study adverse events following immunization (AEFI) with pentavalent vaccine. DESIGN: Cohort study. SETTING: Tertiary-care hospital predominantly catering to urban poor population of East Delhi. PARTICIPANTS: 265 low birthweight (1800-2499 g) and 265 normal birthweight (2500-4000 g) infants. Monovalent Hepatitis B vaccine was administered within 24 hours of birth followed by three primary doses of pentavalent vaccine at 6, 10 and 14 weeks. Anti-HBs titers were estimated after 6 weeks of third dose of pentavalent vaccine. Adverse events following immunization (AEFI) month were observed for a month after each dose of pentavalent vaccine. MAIN OUTCOME MEASURES: Anti HBs antibody titers after 6 weeks of primary immunization, and AEFI. RESULTS: 443 (83.5%) infants (225 low birthweight and 218 normal birthweight infants) completed the follow-up. Seroprotection against hepatitis B virus was achieved in both groups after pentavalent vaccine administration. Anti HBs GMTs in low birthweight infants (194.8 mIU/mL) and normal birthweight infants (204.2 mIU/mL) were comparable (P = 0.17). No serious adverse events were observed in either group. CONCLUSIONS: Three primary doses of pentavalent vaccine administered along with zero dose of Hepatitis B vaccine at birth provide good seroprotection. The vaccine appears to be safe in both low birth weight and normal birthweight infants born at term.

Front loading sputum microscopy - an alternative approach for diagnosis of pulmonary tuberculosis.

INTRODUCTION: Until newer, rapid, economical tools are introduced for diagnosis of Pulmonary Tuberculosis in resource limited settings, optimization of sputum smear examination for increasing case detection remains of utmost priority. The aim of the study was to detect presumptive TB patients using Front Loading sputum microscopy and compare it with Standard method. METHODS: Three sputum specimens (Spot 1- on spot at the time of first visit, Spot 2- one hour after Spot 1 and early morning-next day early morning sample) from 552 TB suspect cases were collected. Zeihl Neelsen staining (spot 1, spot 2 and early morning respectively) and microscopy by Front Loading (spot 1, spot 2) and Standard method (spot 1, early morning) of sputum microscopy were done. RESULTS: Culture on LJ media being the gold standard, the sensitivity and specificity of the Front Loading and the Standard method of sputum microscopy were 68.65%, 94.43% and 70.14%, 93.6% respectively. The difference between two methods was not statistically significant. 91.1% patients gave preference for same day sampling process. CONCLUSION: The sensitivity and specificity of sputum microscopy using an early morning sample followed by another sputum one hour later from the same day appears not to be inferior to using two early morning samples on subsequent days. The Front Loading sputum microscopy can be implemented in DOTS clinic on the day of first visit of patients to health care center to increase compliance of patients with diagnostic procedure and decrease drop-outs.

Clinical and serological characteristics of nail psoriasis in Indian patients: A cross-sectional study.

BACKGROUND: Nail involvement in psoriasis is common with a lifetime incidence of 80-90%. It may reflect severity of cutaneous involvement and predict joint disease. Yet it remains, poorly studied and evaluated especially in Indian psoriatic patients. AIM: The present study was undertaken to evaluate clinical and serological profile of nail involvement in psoriasis and to assess quality of life impairment associated with nail involvement in Indian patients. METHODS: Consecutive patients with nail psoriasis were assessed for severity of cutaneous disease (psoriasis area severity index score) and nail disease (nail psoriasis severity index score). The impairment in quality of life attributable to nail disease was scored with nail psoriasis quality of life 10 score. All patients were also assessed for joint disease and tested for inflammatory and serological markers as erythrocyte sedimentation rate, C-reactive protein, rheumatoid factor and anti-cyclic citrullinated peptide antibodies. RESULTS: In our cohort of 38 patients with nail psoriasis, 9 had concomitant psoriatic arthritis. The mean psoriasis area severity index was 14.4 ± 9.6 (range = 0.4-34). The most commonly recorded psoriatic nail changes were pitting (97.4%), onycholysis (94.7%) and subungual hyperkeratosis (89.5%). The mean nail psoriasis severity index score was 83.2 ± 40.1 (range = 5-156) and mean nail psoriasis quality of life 10 was 1.1 ± 0.4. Erythrocyte sedimentation rate and C-reactive protein were raised in 22/38 (57.9%) and 15/38 (39.5%) patients, respectively; rheumatoid factor was positive in 5/38 (13.2%) and anti-cyclic citrullinated peptide antibody was raised in 4/38 (10.5%) patients. LIMITATIONS: Small sample size and lack of a control group. CONCLUSIONS: In Indian patients with nail psoriasis, severity of nail involvement was found to be poorly correlated with the extent of cutaneous disease. In addition the impact of nail disease on patient's quality of life was found to be minimal. This suggests the need for a quality of life questionnaire suited to the Indian population. Serological markers were raised overall in the study patients and more so in the patients with concomitant arthritis.

Effect of oral cranberry extract (standardized proanthocyanidin-A) in patients with recurrent UTI by pathogenic E. coli: a randomized placebo-controlled clinical research study.

PURPOSE: To evaluate the effect of cranberry extract (PAC-A ~ proanthocyanidin-A) on the in vitro bacterial properties of uropathogenic (E. coli) and its efficacy/tolerability in patients with subclinical or uncomplicated recurrent UTI (r-UTI). MATERIALS AND METHODS: After obtaining clearance from the ethics committee and administering a written informed consent, 72 patients with r-UTI were enrolled as per protocol (November 2011 to March 2013) in this prospective study, to randomly receive (PAC-A: group I, 36) or (placebo: group II, 36), for 12 weeks. Any change/reduction in the incidence of r-UTI at 12 weeks was construed to be the primary endpoint of this study. RESULTS: After 12 weeks, bacterial adhesion scoring decreased (0.28)/(2.14) in group I/II (p < 0.001); 32/36 (88.8 %) and 2/36 (5.5 %) in groups I and II, respectively, turned MRHA negative (p < 0.001); biofilm (p < 0.01) and bacterial growth (p < 0.001) decreased in group I; microscopic pyuria score was 0.36/2.0 in group I/II (p < 0.001); r-UTI decreased to 33.33 versus 88.89 % in group I/II (p < 0.001); mean subjective dysuria score was 0.19 versus 1.47 in group I/II (p < 0.001), while mean urine pH was 5.88 versus 6.30 in group I/II (p < 0.001). No in vitro antibacterial activity of cranberry could be demonstrated, and no adverse events were noted. CONCLUSIONS: The overall efficacy and tolerability of standardized cranberry extract containing (PAC-A) as a food supplement were superior to placebo in terms of reduced bacterial adhesion; bacterial MRHA negativity; urine pH reduction; and in preventing r-UTI (dysuria, bacteriuria and pyuria). Larger randomized controlled trials are needed to elucidate the precise role, exact dose and optimal duration of PAC-A therapy in patients at risk of r-UTI.

Significance of Anti-cyclic Citrullinated Peptide Autoantibodies in Immune-mediated Inflammatory Skin Disorders with and without Arthritis.

BACKGROUND: Anti-cyclic citrullinated peptides (CCPs) are autoantibodies directed against citrullinated peptides. Rheumatoid factor (RF), an antibody against the Fc portion of IgG, is known to form immune complexes and contribute to the etiopathogenesis of various skin disorders. C-reactive protein (CRP), an acute-phase protein, increases following secretion of interleukin-6 from macrophages and T cells. Anti-CCP, RF, and CRP are well-established immune-markers, their diagnostic potential in immune-mediated skin disorders remains less widely studied. AIMS AND OBJECTIVES: To determine the correlation between anti-CCP, RF, and CRP in immune-mediated inflammatory skin diseases. MATERIALS AND METHODS: About 61 clinically diagnosed cases of various immune-mediated skin diseases (psoriasis [n = 38], connective tissue diseases such as systemic lupus erythematosus and systemic sclerosis [n = 14], and immunobullous disorders including pemphigus vulgaris and pemphigus foliaceus [n = 9]) were included in the study. These patients were subclassified on the basis of presence or absence of arthritis. Arthritis was present in nine cases of psoriasis and seven connective tissue disorder patients. Detection of serum anti-CCP was done using enzyme-linked immunosorbent assay, whereas CRP and RF levels were detected using latex agglutination technique. RESULTS: Of the 61 specimens, 14.75% had elevated serum anti-CCP levels. RF and CRP levels were elevated in 18.03% and 39.34% specimens, respectively. RF was elevated in 13.16% of inflammatory and 42.88% of connective tissue disorders, whereas anti-CCP was raised in 10.53% of inflammatory and 35.71% of connective tissue disorders. CRP positivity was highest in connective tissue disorders (50%), followed by 39.47% in inflammatory and 22.22% in immunobullous conditions. In none of the immunobullous patients, anti-CCP or RF levels were found to be elevated. Association of the presence of arthritis with elevated anti-CCP was found to be statistically significant. CONCLUSIONS: Although anti-CCP, RF, and CRP levels are valuable markers of chronic immune-mediated skin disorders, elaborate studies enrolling a larger number of patients are required to validate these diagnostic markers.

Laparoscopy in the diagnosis of tuberculosis in chronic pelvic pain.

BACKGROUND: To estimate the prevalence of genital tuberculosis in women with idiopathic chronic pelvic pain on laparoscopy, correlate laparoscopic findings with microbiological and histological diagnosis of tuberculosis and assess the response to anti tubercular treatment (ATT) in these cases. METHOD: In a prospective cohort study, fifty women with idiopathic chronic pelvic pain were enrolled. Diagnostic laparoscopy was done in all women and fluid from pouch of Douglas and/or saline washings were sent for acid fast bacilli (AFB) smear, conventional and rapid culture and DNA polymerase chain reaction (PCR) analysis for diagnosis of genital TB. The results of these tests were analyzed and agreement with laparoscopy was assessed using Kappa statistics. Pain scores using visual analogue scale were compared before and after treatment. RESULTS: Pelvic pathology was present in 44 (88%) women of idiopathic chronic pelvic pain, with a 34% prevalence rate of genital tuberculosis. Pelvic inflammation was associated with positive peritoneal fluid PCR (n=4) and AFB culture (n=3). Acid fast bacilli PCR had substantial agreement (kappa statistics=0.716) with visual findings at laparoscopy. There was a significant reduction in pain scores after treatment. CONCLUSION: Genital tuberculosis contributes to one-third cases of chronic pelvic pain. Pelvic inflammation is an early feature of genital TB and peritoneal fluid PCR has the best co-relation with laparoscopic findings of genital tuberculosis.

Vitamin D Supplementation for Treatment and Prevention of Pneumonia in Under-five Children: A Randomized Double-blind Placebo Controlled Trial.

OBJECTIVE: To evaluate the efficacy of single oral mega-dose of Vitamin D3 for treatment and prevention of pneumonia in under-five children. DESIGN: Randomized, double blind, placebo-controlled trial. SETTING: Tertiary-care hospital. PARTICIPANTS: 324 children (of 980 assessed) between 6 mo-5 y age (median (IQR): 12 (7,19.8) mo) with WHO-defined severe pneumonia. Of these, 126 (39%) were vitamin D deficient (serum 25(OH)D <12 ng/mL). INTERVENTION: 100,000 IU of oral cholecalciferol (n= 162) or placebo (n= 162) in single dose, administered at enrolment. Outcome variables: Primary: Time to resolution of severe pneumonia and proportion of children having recurrence of pneumonia in next 6 months; Secondary: Change in serum levels of 25(OH)D; immunoglobulins IgA, IgG, IgM, and cathelicidin 2 weeks following supplementation; and time taken for overall resolution of illness. OUTCOME VARIABLES: Primary: Time to resolution of severe pneumonia and proportion of children having recurrence of pneumonia in next 6 months; Secondary: Change in serum levels of 25(OH)D; immunoglobulins IgA, IgG, IgM, and cathelicidin 2 weeks following supplementation; and time taken for overall resolution of illness. RESULTS: Median (95% CI) time for resolution of severe pneumonia was 30 (29, 31) h in the vitamin D group as compared to 31 (29,33) h in the placebo group [adjusted hazard ratio (95% CI): 1.39 (1.11, 1.76); P = 0.005]. The risk of recurrence of pneumonia in next 6 months was comparable in the two groups [placebo: 36/158 (22.8%); vitamin D: 39/156 (25%); RR (95% CI): 1.13 (0.67,1.90); P 0.69]. Proportion of vitamin D deficient children declined from 38% to 4% in the supplementation group, and from 41% to 33% in the placebo group, two weeks after supplementation. There was no significant effect of vitamin D supplementation on serum levels of cathelicidin, IgA and IgG. The time taken for complete recovery from pneumonia, duration of hospitalization, and fever clearance time were comparable for the two groups. No adverse event was noted related to the intervention. CONCLUSION: There is no robust evidence of a definite biological benefit, either for therapy or prevention, to suggest a routine megadose supplement of vitamin D3 for under-five children with severe pneumonia.

Utility of immunochromatographic assay as a rapid point of care test for screening of antenatal syphilis.

BACKGROUND AND OBJECTIVES: Syphilis is one of the most common preventable causes of adverse effects during pregnancy. Antenatal screening prevents the delay between diagnosis and treatment there by reducing the risk of congenital syphilis. The objective of this study was to evaluate the utility of an immunochromatographic assay as a point of care test for antenatal screening of syphilis. MATERIALS AND METHODS: Sera of 200 antenatal mothers were evaluated for serodiagnosis of syphilis by the venereal disease research laboratory (VDRL), Treponema pallidum hemagglutination assay (TPHA) and SD BIOLINE Syphilis 3.0 test. The performance of SD BIOLINE Syphilis 3.0 test was compared with VDRL as screening assay and TPHA as a confirmatory test. RESULTS: The antenatal prevalence of syphilis was found to be 2% by both VDRL and TPHA. The sensitivity, specificity, positive predictive value, and the negative predictive value of SD BIOLINE Syphilis 3.0 test were 75%, 100%, 100%, and 99.45%, respectively. CONCLUSIONS: Antenatal screening and treatment of maternal syphilis are cost-effective health interventions even under the low prevalence of infection. SD BIOLINE Syphilis 3.0 test, although having less sensitivity than the existing testing strategy, can have a tremendous impact on the disease burden if used prudently for the screening of antenatal mothers in peripheral health settings.

Carriage prevalence of carbapenem-resistant Enterobacteriaceae in stool samples: A surveillance study.

BACKGROUND: With more people being exposed to antibiotics, intestinal microflora faces constant pressure of antibiotic selection, which has resulted in the emergence of multidrug resistant strains. This may pose a severe problem as intestinal Enterobacteriaceae members are commonly implicated in human infections. AIMS: This surveillance study was undertaken to investigate the carriage of carbapenem-resistant Enterobacteriaceae (CRE) in the gastrointestinal tract among patients attending the outpatient clinic in a tertiary care center of East Delhi, India. METHOD: We performed a prospective surveillance study to screen 242 Enterobacteriaceae isolates for carbapenemase production from the stool samples of 123 outpatients attending a tertiary care hospital in East Delhi over a four-month period. RESULTS: Twenty-four (9.9 per cent) isolates demonstrated carbapenemase activity among 242 screened Enterobacteriaceae isolates. Four stool samples had two isolates of different species, both eliciting this feature and therefore indicating presence of multiple carbapenem-resistant Enterobacteriaceae (CRE) isolates in a single sample. CONCLUSION: Screening for carriage of CRE in stools of patients undergoing elective or emergency gastrointestinal surgical procedures, with haematological malignancies taking chemotherapy, or those planned for bone marrow transplantation can guide clinicians about gut colonisation of multidrug-resistant Enterobacteriaceae as these groups of patients are at risk of possible endogenous infection.

Immunocytochemical detection of mycobacterial antigen in extrapulmonary tuberculosis.

The aim of the study is to determine whether immunostaining for mycobacterial antigen can contribute to the cytological diagnosis of extrapulmonary tuberculosis (EPTB). The study was carried out on aspirated material of lymph nodes, and other accessible sites, from 65 patients with clinical diagnosis of tuberculosis (TB). Twenty patients, diagnosed by fine-needle aspiration, with non-tuberculous granulomas served as controls. The diagnosis of TB was based on the demonstration of acid-fast bacilli (AFB), culture positivity for Mycobacterium tuberculosis (M. tuberculosis), or response to treatment with standard anti-tubercular therapy. Immunostaining was done using polyclonal antibody to mycobacteria. AFB positivity by Ziehl Neelsen (ZN) staining was 21%, 65.38%, and 68% respectively in Pattern 1 (granulomas alone), in Pattern 2 (granulomas with necrosis), and in Pattern 3 (necrosis alone). Overall AFB positivity was 56.92%. Twenty-eight of 65 cases were negative for AFB on direct smear. Culture was positive in 46% (13/28). Sensitivity and specificity of immunostaining were 96.92% (63/65) and 95%, respectively. Immunoreactivity was seen in 26 (92.8%) of 28 cases which were negative by ZN staining. Except in the case of leprosy, in which cross reactivity was seen, there was no immunoreactivity in the control group. Immunocytochemistry (ICC) had high sensitivity (96.2%) and specificity (95%) in the diagnosis of EPTB. ICC may be a useful adjunct to evaluation of cytomorphology and ZN staining.

Towards the innovation for microbiology curriculum change: students' perception.

For a medical curriculum to be an effective means of learning for today's students, it has to be designed with knowledge of their priorities, needs and abilities. This can be best achieved by inviting students' view-point during curriculum planning. The present study thus elicits opinion of the medical students through a randomly issued set of questionnaires, towards the present microbiology curriculum in order to quantitate from their view-point, the weakness as well as the strengths of the existing curriculum. Their evaluation reveals that they welcome new techniques like problem-based learning but at the same time emphasise the need to integrate what is taught in close association with clinical circumstance. Hence it is important to understand the minds and needs of our students before implementing the syllabus content across to the consumers.

'Test and treat' policy in dyspepsia: time for a reappraisal.

The 'test and treat' policy is the recommended way to eradicate Helicobacter pylori in young patients with uninvestigated dyspepsia if the prevalence of H. pylori is high. India is considered to have a high prevalence of H. pylori. This observational study was conducted in order to discover the prevalence of H. pylori disease in uninvestigated dyspeptic patients, based on stool antigen test and immunoglobulin M (IgM) antibodies in the sera in order to ascertain the role of the 'test and treat' policy in this geographical area. Fifty patients (age <55 years) with uninvestigated dyspepsia presenting to surgery out-patient department of a tertiary teaching hospital in northern India were included in the study. Fifty healthy controls were also included. Dyspeptic patients with alarm symptoms were excluded from the study. Patients and controls were tested for IgM antibodies in their sera and H. pylori antigen in their stools. The positivity of the IgM antibody and H. pylori antigen in stools of the cases was 16% and 8%, respectively, while only 4% IgM positivity was found in the controls' samples. This study highlights the low prevalence of H. pylori in dyspeptic Indian patients. This indicates that initial proton pump therapy may be a better option than the 'test and treat' policy for young patients with uninvestigated dyspepsia.

Clinico-epidemiological profile and high-level aminoglycoside resistance in enterococcal septicemia from a tertiary care hospital in east Delhi.

BACKGROUND: Emergence of high-level aminoglycoside and glycopeptide resistance has significantly contributed to the mortality, particularly in serious enterococcal infections. OBJECTIVES: This study was aimed to determine the prevalence of high-level gentamicin resistance (HLGR), high-level streptomycin resistance (HLSR) and vancomycin resistance in enterococcal isolates recovered from patients with bacteremia. MATERIALS AND METHODS: A total of 110 blood culture isolates of enterococci were recovered from septicemic patients. Routine antibiotic susceptibility testing was performed and screening for ampilcillin, high-level aminoglycoside resistance (HLAR) and high-level vancomycin resistance was done by agar screen method. RESULTS: Out of 110 isolates, Enterococcus faecium accounted for 53% of these isolates, followed by Enterococcus fecalis (33%), Enterococcus casseliflavus (8%), Enterococcus raffinosus (4%) and Enterococcus dispar (2%). Resistance to ampicillin, HLGR, HLSR and HLAR was detected in 58%, 62%, 58% and 54% of the isolates, respectively. No isolate was resistant to vancomycin. CONCLUSION: This study illustrates the high prevalence of HLAR in enterococci from patients with septicemia in our region, which emphasizes the need to predict synergy between beta-lactams and aminoglycosides for management of enterococcal infections.