Multidrug resistance mediated by co-carriage of extended-spectrum beta-lactamases, AmpC and New Delhi metallo-beta-lactamase-1 genes among carbapenem-resistant Enterobacteriaceae at five Indian medical centres.

In this study, we evaluated the coexistence of extended-spectrum beta-lactamases (ESBL), AmpC and New Delhi metallo-beta-lactamase-1 (NDM-1) genes among carbapenem-resistant Enterobacteriaceae (CRE) recovered prospectively from patients at multiple sites. The study included 285 CRE strains from 2782 Gram-negative Bacilli collected from multiple centres during 2007-2010, of which 87 were characterised. Standard and reference laboratory methods were used for resistance determination. Detection of blaNDM-1 , blaAmpC , blaTEM , blaSHV and blaCTX-M was done by polymerase chain reaction. High levels of antimicrobial resistance observed among study isolates. Co-carriage of ESBLs, AmpC and NDM-1 was 26.3%. Nosocomial origin among the co-carriage isolates was 64.3%, with 9.2% associated mortality.

Assessing effect of climate on the incidence of dengue in Tamil Nadu.

Incidence of dengue is reported to be influenced by climatic factors. The objective of this study is to assess the association of local climate with dengue incidence, in two geographically distinct districts in Tamil Nadu. The study uses climate data, rainfall and mean maximum and minimum temperature to assess its association if any, with dengue incidence in two districts of Tamil Nadu, South India. According to this study while precipitation levels have an effect on dengue incidence in Tamil Nadu, non-climatic factors such as presence of breeding sites, vector control and surveillance are important issues that need to be addressed.

The Chennai Declaration: a roadmap to tackle the challenge of antimicrobial resistance.

"A Roadmap to Tackle the Challenge of Antimicrobial Resistance - A Joint meeting of Medical Societies in India" was organized as a pre-conference symposium of the 2 nd annual conference of the Clinical Infectious Disease Society (CIDSCON 2012) at Chennai on 24 th August. This was the first ever meeting of medical societies in India on issue of tackling resistance, with a plan to formulate a road map to tackle the global challenge of antimicrobial resistance from the Indian perspective. We had representatives from most medical societies in India, eminent policy makers from both central and state governments, representatives of World Health Organization, National Accreditation Board of Hospitals, Medical Council of India, Drug Controller General of India, and Indian Council of Medical Research along with well-known dignitaries in the Indian medical field. The meeting was attended by a large gathering of health care professionals. The meeting consisted of plenary and interactive discussion sessions designed to seek experience and views from a large range of health care professionals and included six international experts who shared action plans in their respective regions. The intention was to gain a broad consensus and range of opinions to guide formation of the road map. The ethos of the meeting was very much not to look back but rather to look forward and make joint efforts to tackle the menace of antibiotic resistance. The Chennai Declaration will be submitted to all stake holders.

Does solid culture for tuberculosis influence clinical decision making in India?

SETTING: Medical units at an academic tertiary referral hospital in Southern India. OBJECTIVE: To investigate the impact of solid culture on Löwenstein-Jensen medium on clinical decision making. DESIGN: In a retrospective review of 150 culture-positive and 150 culture-negative consecutively sampled tuberculosis (TB) suspects, treatment decisions were analysed at presentation, after the availability of culture detection results and after the availability of drug susceptibility testing (DST) culture results. RESULTS: A total of 124 (82.7%) culture-positive patients and 35 (23.3%) culture-negative patients started anti-tuberculosis treatment prior to receiving their culture results; 101 patients (33.7%) returned for their results; two (1.3%) initiated treatment based on positive culture and no culture-negative patients discontinued treatment. DST was performed on 119 (79.3%) positive cultures: 30 (25.2%) showed any resistance, eight (6.7%) showed multidrug resistance and one (0.84%) showed extensively drug-resistant TB. Twenty-eight patients (23.5%) returned for their DST results. Based on DST, treatment was modified in four patients (3.4%). CONCLUSION: Using solid culture, 150 cultures need to be tested for one treatment modification and 30 for DST. The cost of the widespread application of culture will need to be balanced against its impact on treatment decisions in India.

Correlation of TEM, SHV and CTX-M extended-spectrum beta lactamases among Enterobacteriaceae with their in vitro antimicrobial susceptibility.

PURPOSE: The present study was carried out to characterize the ESBL types and evaluated their in vitro activity against a collection of Gram negative bacteria (GNB) from a multicentric Indian surveillance study. MATERIAL AND METHODS: During January 2005 to June 2006, six tertiary care centres in India forwarded 778 non-duplicate GNB to our reference laboratory. Three hundred GNB from this collection were selected based on clinical significance and were used in the present study. Tested isolates included Escherichia coli (167), Klebsiella spp. (122) and Enterobacter spp. (11). ESBL screening and confirmation was performed for all the isolates. Minimum inhibitory concentration of imipenem, meropenem, ertapenem, levofloxacin, amikacin, piperacillin/tazobactam and ceftriaxone was determined by the E-test method. Molecular typing of the ESBLs was performed by polymerase chain reaction among the 121 selected isolates. RESULTS: The study showed excellent susceptibility among the strains to imipenem (100%), meropenem (100%) and ertapenem (98.7%); good susceptibility to amikacin (89.7%) and piperacillin/tazobactam (85.3%) was observed. TEM and CTX-M were predominantly found in E. coli (39.2%) while, among the Klebsiella spp., TEM, SHV and CTX-M occurred together in 42.6% of the isolates. CONCLUSION: More than one ESBL was produced by many strains, and this was correlated with increased resistance levels. Carbapenems continue to show good in vitro activity and ertapenem is a potential alternative to imipenem and meropenem. Continued antimicrobial resistance surveillance is warranted in light of these findings.

Detection of human immunodeficiency virus infection in the sputum of tuberculosis patients in South India.

SETTING: A DOTS clinic in an academic tertiary referral hospital in South India. OBJECTIVE: To evaluate the performance of two rapid enzyme-linked immunoassays (EIAs) for the detection of human immunodeficiency virus (HIV) infection in sputum samples of patients with tuberculosis (TB). DESIGN: We prospectively recruited 522 consecutive out-patients presenting to the DOTS clinic with confirmed TB of any type to undergo HIV testing using reference serum EIA and index-blinded parallel sputum HIV testing with two rapid EIAs designed for oral mucosal transudate. RESULTS: HIV positivity was 14.9% (95%CI 12.1-18.4). Compared to reference serum EIA, the Oraquick™ assay was 93.1% sensitive (95%CI 83.8-97.4) and 95.3% specific (95%CI 92.7-96.9), while the Aware™ assay was 92.3% sensitive (95%CI 83.4-96.8) and 96.6% specific (95%CI 94.4-98.0). The positive predictive values were respectively 77.0% and 82.7%. After freezing of sputum, the sensitivity of both assays declined, but the specificity significantly increased. Higher sputum volume reduced the odds of obtaining a true result with both assays. CONCLUSION: HIV testing of fresh sputum is not sufficiently accurate for anonymous HIV surveillance among TB patients in a setting of low (<10%) HIV prevalence. Freezing sputum samples and limiting sputum volume for HIV testing may improve assay specificity.

Detection and characterization of metallo beta lactamases producing Pseudomonas aeruginosa.

This study was undertaken to evaluate phenotypic and genotypic methods for detection of Metallo-Beta-Lactamases (MBLs) among nosocomial Pseudomonas aeruginosa. Sixty one among 176 P. aeruginosa isolates, collected as part of a multicentric study (2005-2007), were evaluated for carbapenem resistance (CARB-R; resistant to either imipenem/meropenem) and screened for MBL by Combination Disk Diffusion Test (CDDT) using imipenem (IMP), meropenem (MER) and ceftazidime (CAZ) with EDTA. MBL positives were further confirmed by IMP + EDTA Etest. Twenty strains (42.6%) were found to be MBL producers among the 61 P. aeruginosa. PCR for IMP and VIM MBL was performed on 48 of the 61, 15 were positive for VIM MBL type. CDDT using IMP + EDTA had the highest sensitivity and specificity of 87.8% and 84.4% when compared to Etest, which was higher than the values obtained for CAZ + EDTA and MER + EDTA. CDDT using IMP + EDTA also compared very well with the PCR (specificity = 90.9%, sensitivity = 93.3%). CARB-R among P. aeruginosa is mediated predominantly via MBL production. Clinical P. aeruginosa isolates can be screened routinely using the less expensive IMP + EDTA CDDT in clinical microbiology laboratories.

Liposomal amphotericin B (Fungisome) for the treatment of cryptococcal meningitis in HIV/AIDS patients in India: a multicentric, randomized controlled trial.

BACKGROUND: There is need to investigate the use of liposomal amphotericin B in cryptococcal meningitis in India. AIMS: To compare the efficacy, safety, duration of treatment and cost of two doses of liposomal amphotericin B (Amp B) (Fungisome) in cryptococcal meningitis in HIV/AIDS patients. SETTINGS AND DESIGN: Prospective, randomized, multicenter study in tertiary care hospitals across India. MATERIALS AND METHODS: Adult patients with culture-proven cryptococcal meningitis with HIV/AIDS were randomized to receive either 1 (Group A) or 3 mg/kg/day of Fungisome (Group B). Clinical efficacy and tolerability, laboratory evaluations and mycological response were assessed daily, twice weekly and weekly respectively. The patients were assessed at four and eight-week follow-up. STATISTICS: We calculated average and standard deviation for the various parameters. RESULTS: The time to show clinical response was 13.66 days (1 mg) and 9.55 days (3 mg). In Group B (n=6 complete response), 50% patients responded within one week by microbial conversion, 83% in two weeks and 100% in three weeks. Patients with 1 mg dose (n=4 complete response), none showed microbial conversion within one week, 75% responded in two weeks, whereas one patient took four weeks. The average duration of treatment was 36.5+/-14.4 and 26.5+/-5.89 (S.D.) days in 1 and 3 mg/kg/day respectively. Drug was tolerated with little renal, hepatic or hematological toxicity. The cost was found to be 3.81 lacs and 1.74 lacs with 3mg/kg/day and 1mg/kg/day respectively. CONCLUSION: Higher dose showed better efficacy and quicker microbial conversion of Cerebrospinal fluid (CSF) (cerebrospinal fluid) than 1 mg/kg/day. It shortened the duration of treatment in days by 27% while drug cost almost doubled ( CLINICAL TRIAL REGISTRATION NUMBER: ISRCTN 52812742).

Diagnostic accuracy of the microscopic observation drug susceptibility assay: a pilot study from India.

SETTING: The microscopic observation drug susceptibility (MODS) assay is a rapid, sensitive, low-cost liquid culture technique. OBJECTIVE: To establish the accuracy of MODS for the detection of active pulmonary tuberculosis (TB), and to document the costs and challenges of setting up this assay in a low-income setting. DESIGN: Prospective blinded pilot study of 200 adult TB suspects at a tertiary referral hospital in India. Reference standard included culture (Löwenstein-Jensen and automated liquid culture) and clinical diagnosis. RESULTS: Patients were mostly male (n = 122, 61.1%) and out-patients (n = 184, 92.0%), with a mean age of 40.4 years (standard deviation 16.2). Seventeen (8.5%) were human immunodeficiency virus infected and 47 (23.5%) were reference culture-positive. Compared to reference culture, MODS was 78.9% sensitive (95%CI 62.2-90.0) and 96.7% specific (95%CI 92.0-98.8). Clinical assessment suggested that MODS was false-negative in 3/8 reference culture-positive MODS-negatives and true-positive in 4/6 reference culture-negative MODS-positives. MODS was faster than solid (P < 0.001) and liquid culture (P = 0.088), and cheaper than both. CONCLUSION: MODS may be a good alternative to automated liquid culture, but there were several challenges in setting up the assay. Prior training and validation, setup costs and inability to rule out cross-contamination need to be taken into account before the test can be established.

Blinded evaluation of commercial urinary lipoarabinomannan for active tuberculosis: a pilot study.

SETTING: Urine antigen testing is an attractive strategy for the diagnosis of active tuberculosis (TB), but accuracy data are scarce. OBJECTIVE: To prospectively evaluate the diagnostic performance of commercial urinary lipoarabinomannan (LAM) antigen testing for active TB among pulmonary and extra-pulmonary TB suspects. DESIGN: Prospective blinded evaluation of 200 adult TB suspects at a tertiary referral hospital in India. Reference standards included culture and clinical diagnosis. RESULTS: Patients were 61% male (mean age 40.4 years): 8.5% were human immunodeficiency virus (HIV) infected and 47 of 200 (23.5%) were culture-positive for TB. Compared to positivity on either Löwenstein-Jensen (LJ) or BACTEC cultures, LAM sensitivity was 17.8% (95%CI 8.5-32.6), while specificity was 87.7% (95%CI 81.3-92.3). Compared to positivity on both LJ and BACTEC, LAM sensitivity was 5.8% (95%CI 12.5-44.9), with a specificity of 88.8% (95%CI 82.7-92.9). Compared to the clinical diagnosis, LAM sensitivity was 20.0% (95%CI 1.1-70.1), with a specificity of 83.3% (95%CI 50.9-97.0). HIV and smear status did not influence test accuracy. CONCLUSION: In its current form, LAM is insensitive for the diagnosis of active TB, although its specificity is adequate.

Granulomatous hepatitis following open heart surgery with cardiopulmonary bypass.

BACKGROUND: Cardiopulmonary bypass (CPB) initiates an inflammatory cascade, predisposing the patient to a number of infections. The stress of surgery and anaesthesia further expose the patient to a variety of non-infectious complications. We report a group of patients who developed granulomatous disease after open heart surgery. METHODS: We retrospectively analysed a subset of patients who developed a syndrome of fever, jaundice and hepatomegaly after open heart surgery. We recruited age- and sex-matched controls who underwent open heart surgery during the same period (July 2002-July 2004). Details of demographic profiles, diagnostic evaluation and drug treatment were noted and compared between the two groups using the SPSS software. RESULTS: Five patients were identified to have the specific syndrome of high grade intermittent fever with jaundice and hepatomegaly with investigations revealing an intrahepatic cholestasis. A detailed evaluation revealed granulomas in tissue specimens of the bone marrow and/or liver in these patients. An extensive evaluation for an alternative aetiological agent was non-contributory. CONCLUSION: We found granulomatous hepatitis in 5 patients following open heart surgery and they were given conventional antituberculous therapy to which they responded. It is possible that in these patients, tuberculosis was re-activated from a dormant focus due to a period of transient immunodeficiency caused by an extracorporeal circulation.

Scrub typhus among hospitalised patients with febrile illness in South India: magnitude and clinical predictors.

OBJECTIVES: To derive a clinical algorithm for diagnosis of scrub typhus among patients hospitalized with febrile illness and to determine predictors of bad prognosis. METHODS: Patients hospitalized with febrile illness of 5-30 days duration were evaluated for common aetiologies. Sera were tested for antibodies (IgM and IgG) to Orientia tsutsugamushi using ELISA kit. RESULTS: Among 207 patients, 50 had elevated levels of IgM antibodies. The data of these patients were compared with that of 16 controls having febrile illnesses in whom rickettsial infection was ruled out. Transaminase elevation (>twice normal) was present in 90% and was significantly (P=0.004) more common in those with scrub typhus. If a combination of elevated transaminases, thrombocytopenia and leukocytosis is used, the specificity and positive predictive value are about 80%. Case fatality rate was 14%. Univariate analysis showed that hyperbilirubinemia (>1.5mg%) has a RR of 9 (95% CI=1.48-58.5) and elevated creatinine level (>1.4 mg%) had a RR of 43.99 (95% CI=3.65-530.5) for death. Elevated creatinine level was found to be an independent predictor of mortality (P=0.02). CONCLUSION: In developing countries with limited diagnostic facilities, it is prudent to recommend empiric therapy in patients with undifferentiated febrile illness having evidence of multiple system involvement especially if there is transaminase elevation. Elevated creatinine may predict bad outcome.

Disseminated histoplasmosis.

OBJECTIVE: To study the clinical features and natural history of disseminated histoplasmosis(DH) in India. METHODS: We retrospectively analyzed the data obtained from the in-patient medical records of adults (age > 13 years) diagnosed to have DH during the period from January 1989 to December 1999. DH was diagnosed when histologically compatible intracellular organisms were present or Histoplasma capsulatum was obtained in culture from the extrapulmonary sites. RESULTS: Nineteen patients (18 male and 1 female) were diagnosed to have DH. Diabetes mellitus and HIV infection were the most common co-morbid conditions. Weight loss, fever and oropharyngeal ulcers were the commonest symptoms. Physical signs included hepatosplenomegaly, oropharyngeal ulcers and lymphadenopathy. The diagnosis was confirmed by histopathology and/or culture from the following sites: bone marrow, adrenal gland, lymph node, oropharyngeal ulcers, rectal mucosa and skin. Two patients were treated with Amphotericin B, 6 with various azoles and 3 had Amphotericin B followed by various azoles. Among the eleven treated, 7 were cured, 2 improved, 1 had a relapse and 1 patient died. CONCLUSION: DH is not uncommon in India and should be considered in the diagnosis of patients with prolonged fever, weight loss, oropharyngeal ulcers, hepatosplenomegaly, lymphadenopathy and adrenal enlargement. Correct diagnosis and treatment leads to a favourable outcome.

Subtype & cytokine profiles of HIV infected individuals from south India.

BACKGROUND & OBJECTIVE: The global surveillance of human immunodeficiency virus (HIV) subtypes (clades) helps understand the global distribution and incidence of different HIV subtypes. As knowledge about subtypes circulating in an area is needed for developing a candidate vaccine, prevalence of the subtypes HIV-1 and HIV-2 were studied in south India. The profile of cytokines interleukin 10 (IL10) and interferon gamma (IFNgamma) in both types of infection were also analysed as these are considered indicators of disease progression. METHODS: Patients who belonged to the 4 south Indian States i.e. Tamil Nadu, Kerala, Karnataka and Andhra Pradesh were included. HIV-1 subtyping was carried out by the heteroduplex mobility analysis (HMA) while that of HIV-2 was done by direct sequencing. The quantitation of IFNgamma and IL-10 was carried out using commercial ELISA kits. RESULTS: Among the 82 HIV-1 infected individuals subtyped, 78 (95.1%) were subtype C while all 12 HIV-2 strains were subtype A. IL-10 concentration was significantly higher among HIV infected individuals compared to normal healthy controls. IFNgamma was significantly higher among symptomatic and AIDS groups compared to asymptomatic HIV-1 infected individuals. INTERPRETATION & CONCLUSION: HIV-1 subtype C and the HIV-2 subtype A are the major subtypes circulating in south India. The study showed a trend towards a shifting of the cytokine profile from Th1 to Th2/Th0 in HIV-1, HIV-2 infections, and HIV-1 and HIV-2 dual infected individuals as the disease progresses. This trend observed is not unlike that reported from the West, despite the difference in subtype profile.

Hyperglycaemia as an indicator of concurrent acute pancreatitis in fulminant hepatic failure associated with hepatitis B infection.

Pancreatitis occurring concurrently with fulminant hepatic failure (FHF) is primarily detected on autopsy and is seldom clinically apparent. We report a fatal case of FHF in a 25-year-old woman which was related to acute hepatitis B infection. In this patient, hyperglycaemia needing insulin infusions led to the detection of acute pancreatitis. FHF complicated by acute pancreatitis has a poor prognosis. A high index of suspicion is necessary for its diagnosis. The role of orthotopic liver transplantation and use of antiviral therapies need further evaluation in this situation.

Varied clinical manifestations of Penicillium marneffei in patients with human immunodeficiency virus: a report from south India.

Penicillium marneffei is a dimorphic fungus endemic to northeast India. This is the first report of penicilliosis from a hospital in south India. Three patients with diverse clinical manifestations who presented to a tertiary care hospital are described. High index of clinical suspicion and knowledge of 'local' microbiology is paramount in early diagnosis and timely treatment.

Predictors of multi-organ dysfunction in heatstroke.

BACKGROUND: Heatstroke is a medical emergency that results from failure of thermoregulatory mechanism coupled with an exaggerated acute phase response, causing an elevation in core body temperature that rises above 40 degrees C, producing multi-organ dysfunction. It carries a high mortality rate, and in survivors, a risk of permanent neurological damage. OBJECTIVE: To investigate predictors of multiple organ dysfunction syndrome in patients presenting with heatstroke. METHODS: We investigated 28 patients admitted to a hospital in southern India during the period January 1998 to December 2001. Using a standard form, we collected data on the patients' characteristics, laboratory data, and outcome, and compared those with multiple organ dysfunction with those without such dysfunction. RESULTS: We found that more than three quarters of the studied patients developed multiple organ dysfunction, with the most common dysfunction being respiratory failure. Among the selected predictors, metabolic acidosis 14 of 16 patients, 87.5%; p = 0.011, elevated CPK 17 of 19 patients, 89.5%; p = 0.005, and liver enzymes elevated more than twice the normal (11 of 18 patients, 61%; p = 0.02) had the highest correlation with dysfunction of two or more organs. CONCLUSIONS: The high mortality observed in heatstroke is secondary to multi-organ dysfunction, and among the various parameters assessed, high levels of CPK (>1000 IU/l), metabolic acidosis, and elevated liver enzymes are predictive. Aggressive measures to lower the body temperature with other supportive therapy could substantially reduce the mortality.

Clinical manifestations and management of cryptococcal infection.

Cryptococcus neoformans is an important fungal pathogen causing invasive infection, especially of the central nervous system in this era of the HIV/AIDS epidemic. The choice of treatment depends on site(s) of infection and the patient's immune status. Use of appropriate antifungal agents decreases mortality significantly, but requires continued therapy and long-term maintenance to prevent relapses. The use of liposomal amphotericin B (L Amp B) has overcome some of the difficulties usually found in this setting. The major advantage of these liposomal formulations are faster clearance of C. neoformans [cerebrospinal fluid (CSF) negative] and a reduction in amphotericin toxicity. The majority of clinical efficacy data related to L Amp B are derived from compassionate use studies and case series. Use of liposomal amphotericin has also shown to be a cost effective approach.

The efficacies of affected-limb care with penicillin diethylcarbamazine, the combination of both drugs or antibiotic ointment, in the prevention of acute adenolymphangitis during bancroftian filariasis.

Repeated attacks of adenolymphangitis (ADL) contribute significantly to the progression of chronic lymphoedema in lymphatic filariasis. They are a cause of stigma and, since they may prevent work and require treatment for which payment must be made, of economic loss. The aim of the present study was to improve the treatment of ADL attacks, which is currently mostly empirical. In a double-blind, placebo-controlled, clinical study, 150 subjects who had each suffered at least two ADL attacks in the preceding year were enrolled and randomly allocated to a programme of self-care of the affected limb (after an intensive training programme) and one of five treatments for 12 months. The subjects were supplied with tablets and ointment so that they could take oral penicillin (800 mg/day), oral diethylcarbamazine (DEC; 1 mg/kg.day) or both of these drugs (at the same doses), or apply framycetin ointment to the affected limb, or just take placebo tablets and apply placebo (zinc-oxide) ointment. Placebo tablets and placebo ointment were used so that neither the subjects nor those assessing the responses to treatment were aware of the treatment arm to which each subject had been assigned. The subjects were requested to continue with the affected-limb care after they had stopped taking the tablets and applying the cream, and were followed-up for 24 months from the first treatment.Overall, the mean incidence of ADL attacks decreased from 2.7 episodes/person-year in the pre-treatment year to just 0.38 episode/person-year during the treatment year (P< 0.01). The greatest reduction in incidence was seen in the 58 subjects who received penicillin (with or without DEC). Even in the placebo group, however, the incidence of ADL in the treatment year was significantly lower than that seen in the pre-treatment year, indicating that affected-limb care on its own helps to prevent some attacks. In all groups except the placebo, the incidence of ADL attacks in the year post-treatment exceeded that seen in the treatment year, indicating that chemoprophylaxis needs to be continued for more than a year if such attacks are to be prevented. In most (84%) of the attacks recorded, titres of anti-streptococcal antibodies were seen to be elevated (compared with those recorded during convalescence),indicating that streptococci have a role in the aetiology of ADL. It is recommended that a combination of penicillin prophylaxis and affected-limb care be incorporated into filariasis-control programmes, to decrease morbidity.