Procalcitonin as a prognostic marker for sepsis: a prospective observational study.

BACKGROUND: Procalcitonin is useful for the diagnosis of sepsis but its prognostic value regarding mortality is unclear. This prospective observational study was designed to study the prognostic value of procalcitonin in prediction of 28 day mortality in patients of sepsis. Fifty-four consecutive patients of sepsis, severe sepsis and septic shock defined using the 2001 Consensus Conference SCCM/ESICM/ACCP/ATS/SIS criteria from medical Intensive Care Unit (ICU) of a tertiary care center in New Delhi, India were enrolled from July 2011 to June 2013. Procalcitonin (PCT), C-reactive protein (CRP) measurements were recorded on day 1, day 7 and day 28 of follow up. RESULTS: Procalcitonin value was a better predictor of all-cause short-term mortality than C-reactive protein. Those patients with Procalcitonin levels <7 ng/ml showed higher cumulative survival than those with level [greater than or equal to]7 ng/ml (69.1% vs. 39.5%, p = 0.02). No such effect was observed in relation to C-reactive protein. Procalcitonin levels [greater than or equal to]7 ng/ml predicted mortality with a hazard ratio of 2.6(1.1-6.3). CONCLUSIONS: A Procalcitonin value [greater than or equal to]7 ng/ml obtained at the time of admission to the ICU is a predictor of short-term mortality and thus may allow the identification of those septic patients at increased mortality risk, and help improve their treatment.

Comparative evaluation of two flowcytometric analysers as diagnostic tools for the automated detection of malaria.

GOALS: Automated flowcytometric analysers are used to analyse complete blood count (CBC) in most patients. They differentiate WBC depending on their size, internal granularity, and nuclear content by producing various scattergrams. In malaria, haemozoin pigment-containing cells (parasitized RBC and phagocytic cells) are able to depolarize the laser beam and therefore exhibit abnormal scattergram. PROCEDURE: A study was conducted to evaluate the diagnostic utility and to determine the sensitivity and specificity of flowcytometric haematology analysers [Sysmex XE-2100 (System 1) and Cell-Dyn 3700 (System 2)] in comparison with the conventional microscopy (QBC, AO, Giemsa) and ICT (Immunochromatographic test) for detection of the malaria parasite. RESULTS: Fifty-eight of 320 samples were found to be positive for malaria by conventional methods. Considering atypical scattergram, System 1 showed the sensitivity and specificity of 68.9% and 90.6%, respectively; whereas System 2 showed a sensitivity of 100% and specificity of 98.8%. CONCLUSION: System 2 (Cell-Dyn 3700) was found to be a highly sensitive and specific tool compared to System 1 and conventional methods. Hence, it may be preferred for automated detection of malaria in the blood samples of patients with a differential diagnosis of malaria and those who are subjected to CBC analysis.

Clostridium difficile infections in HIV-positive patients with diarrhoea.

BACKGROUND: Patients with HIV/AIDS are at a high risk of being infected with toxin-producing strains of Clostridium difficile (C. difficile) because of frequent hospitalization, exposure to antibiotics and antibiotic prophylaxis for opportunistic infections. There are little data from India on the prevalence of C. difficile infection in such patients. METHODS: We assessed the occurrence of C. difficile infections in HIV-positive patients with diarrhoea by looking for the presence of its toxin as well as by culturing. Enzyme immunoassay (EIA, Premier toxins A and B; Meridian Diagnostic Inc.) was used to detect toxin from 237 fresh stool samples collected from HIV-positive patients with diarrhoea. Culture was done on cycloserine-cefoxitin-fructose agar and brain- heart infusion agar. RESULTS: C. difficile was found in 12 of 237 (5.1%, 95% CI 2.64%-8.68%) HIV-positive patients with diarrhoea (9 patients were positive by EIA and 3 by culture). The presence of C. difficile in patients who had received antiretroviral therapy (7/66 [10.6%]) was significantly higher (p < 0.016) compared with those who had not (5/171 [3%]). Of the 12 patients positive for C. difficile, 7 were on antiretroviral therapy for a mean (SD) of 34.4 months with mean CD4+ count of 186 (98.81) cells/cmm and 5 patients were anti-retroviral-naïve with mean CD4+ count of 181 (68.7) cells/cmm. All the 12 patients were on antibiotics for previous 2 months and 4 of 12 had been hospitalized in the previous 30 days. CONCLUSION: C. difficile infections occurred more frequently in patients who had received antiretroviral therapy. Our study population had a lower frequency of C. difficile infections compared to previous studies.

Rapid identification of clinical mycobacterial isolates by protein profiling using matrix assisted laser desorption ionization-time of flight mass spectrometry.

PURPOSE: The purpose of this study was to evaluate the identification of Mycobacterium tuberculosis which is often plagued with ambiguity. It is a time consuming process requiring 4-8 weeks after culture positivity, thereby delaying therapeutic intervention. For a successful treatment and disease management, timely diagnosis is imperative. We evaluated a rapid, proteomic based technique for identification of clinical mycobacterial isolates by protein profiling using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). MATERIALS AND METHODS: Freshly grown mycobacterial isolates were used. Acetonitrile/trifluoroacetic acid extraction procedure was carried out, following which cinnamic acid charged plates were subjected to identification by MALDI-TOF MS. RESULTS: A comparative analysis of 42 clinical mycobacterial isolates using the MALDI-TOF MS and conventional techniques was carried out. Among these, 97.61% were found to corroborate with the standard methods at genus level and 85.36% were accurate till the species level. One out of 42 was not in accord with the conventional assays because MALDI-TOF MS established it as Mycobacterium tuberculosis (log (score)>2.0) and conventional methods established it to be non-tuberculous Mycobacterium. CONCLUSIONS: MALDI-TOF MS was found to be an accurate, rapid, cost effective and robust system for identification of mycobacterial species. This innovative approach holds promise for early therapeutic intervention leading to better patient care.

Thrombotic microangiopathy and acute kidney injury following vivax malaria.

BACKGROUND: Infection with Plasmodium vivax, a common human parasite, is occasionally recognized to cause severe organ dysfunction similar to P. falciparum infection. Acute kidney injury (AKI) in malaria is attributed to acute tubular necrosis; thrombotic microangiopathy is not described. METHODS: This observational study includes patients referred to a tertiary care center in North India during June to September 2011 with severe AKI, anemia, and thrombocytopenia following vivax malaria. Renal biopsies were processed by light, immunofluorescence, and electron microscopy. RESULTS: Nine patients (including 5 children) had persistent AKI with thrombocytopenia and variable anemia following the diagnosis of malaria. Based on peripheral smear, eight patients were diagnosed with vivax malaria and had received antimalarial therapy prior to referral; a laboratory diagnosis of P. vivax infection was made for one patient at this center. Renal histology in all cases showed features of thrombotic microangiopathy, including fibrin thrombi, subendothelial widening, and mesangiolysis, along with variable tubulointerstitial nephritis and acute tubular or cortical necrosis. Ultrastructural examination confirmed endothelial injury and subendothelial widening. All patients required hemodialysis, and six were dialysis dependent at four weeks. Delayed presentation to the hospital (P = 0.019), hemolysis on peripheral smear (P = 0.083), and prolonged oligoanuria (P = 0.036) were associated with dialysis dependence. CONCLUSION: The association of anemia, thrombocytopenia, and renal histological evidence of thrombotic microangiopathy with vivax malaria is novel, and suggests the presence of severe endothelial injury. Further studies are necessary to confirm the association and examine the factors associated with its occurrence.

Human bertiellosis from north India.

A 4-y-old boy presented with a history of episodes of gripping abdominal pain and intermittent shedding of segments of a worm in stool, which used to actively change in size and shape. On the basis of measurements of the segments and characteristic features of the eggs, the worm was identified as Bertiella studeri. The characteristic morphological details of the worm, the pitfalls in its identification and the zoonotic potential of this rare cestodal infection are discussed here.

A rare case of cutaneous larva migrans due to Gnathostoma sp.

A 28-year-old lady presented with recurrent erythematous skin lesions in different parts of the body for 3 months. There were several episodes of worm coming out of the lesions. Examination of the worms in the parasitology laboratory revealed it to be a larva of Gnathostoma sp. She was advised treatment with Albendazole for 21 days, and there was no recurrence of lesions.

Early versus delayed initiation of antiretroviral therapy for Indian HIV-Infected individuals with tuberculosis on antituberculosis treatment.

BACKGROUND: For antiretroviral therapy (ART) naive human immunodeficiency virus (HIV) infected adults suffering from tuberculosis (TB), there is uncertainty about the optimal time to initiate highly active antiretroviral therapy (HAART) after starting antituberculosis treatment (ATT), in order to minimize mortality, HIV disease progression, and adverse events. METHODS: In a randomized, open label trial at All India Institute of Medical Sciences, New Delhi, India, eligible HIV positive individuals with a diagnosis of TB were randomly assigned to receive HAART after 2-4 or 8-12 weeks of starting ATT, and were followed for 12 months after HAART initiation. Participants received directly observed therapy short course (DOTS) for TB, and an antiretroviral regimen comprising stavudine or zidovudine, lamivudine, and efavirenz. Primary end points were death from any cause, and progression of HIV disease marked by failure of ART. FINDINGS: A total of 150 patients with HIV and TB were initiated on HAART: 88 received it after 2-4 weeks (early ART) and 62 after 8-12 weeks (delayed ART) of starting ATT. There was no significant difference in mortality between the groups after the introduction of HAART. However, incidence of ART failure was 31% in delayed versus 16% in early ART arm (p = 0.045). Kaplan Meier disease progression free survival at 12 months was 79% for early versus 64% for the delayed ART arm (p = 0.05). Rates of adverse events were similar. INTERPRETATION: Early initiation of HAART for patients with HIV and TB significantly decreases incidence of HIV disease progression and has good tolerability. TRIAL REGISTRATION: CTRI/2011/12/002260.

Prevalence of HIV drug resistance mutation in the northern Indian population after failure of the first line antiretroviral therapy.

There is limited information available about the prevalence and pattern of human immunodeficiency virus (HIV) drug resistance mutations (DRMs) among antiretroviral therapy (ART) experienced patients from northern India. Results of genotypic drug resistance testing were obtained from plasma samples of 128 patients, who had presented with clinical or immunological failure to treatment after at least six months of ART. Major DRMs associated with any of the three classes of antiretroviral (ARV) drugs, nucleoside reverse transcriptase inhibitors (NRTI), non-nucleoside reverse transcriptase inhibitors (NNRTI) and protease inhibitors (PI), were seen in 120 out of 128 patients (93.8% prevalence). NRTI and NNRTI DRMs were each seen in 115/128 (89.8%) patients, with M184V, M41L, D67N and T215Y being the most frequent among NRTI associated mutations, and K103N, G190A, Y181C and A98G among NNRTI associated ones. PI DRMs were observed in 14/128 (10.9%) patients, with L10I, V82A and L89V being the commonest. These results present a high prevalence of DRMs among ART experienced patients from northern India with clinical or immunological failure of therapy. It emphasizes the need for regular testing of plasma samples of such patients for DRMs in order to detect and replace a failing regimen early, and also the use of HIV drug resistance genotyping of ART naive individuals prior to initiating first line ART for possible transmitted resistance. It is very important to enhance the access of patients to ARV drugs so that their compliance could be improved and hence development of DRMs be minimized.

Prevalence of HIV Drug Resistance Mutations in HIV Type 1 Isolates in Antiretroviral Therapy Naïve Population from Northern India.

Objective. The increased use of antiretroviral therapy (ART) has reduced the morbidity and mortality associated with HIV, adversely leading to the emergence of HIV drug resistance (HIVDR). In this study we aim to evaluate the prevalence of HIVDR mutations in ART-naive HIV-1 infected patients from northern India. Design. Analysis was performed using Viroseq genotyping system based on sequencing of entire protease and two-thirds of the Reverse Transcriptase (RT) region of pol gene. Results. Seventy three chronic HIV-1 infected ART naïve patients eligible for first line ART were enrolled from April 2006 to August 2008. In 68 patients DNA was successfully amplified and sequencing was done. 97% of HIV-1 strains belonged to subtype C, and one each to subtype A1 and subtype B. The overall prevalence of primary DRMs was 2.9% [2/68, 95% confidence interval (CI), 0.3%-10.2%]. One patient had a major RT mutation M184V, known to confer resistance to lamivudine, and another had a major protease inhibitor (PI) mutation D30N that imparts resistance to nelfinavir. Conclusion. Our study shows that primary HIVDR mutations have a prevalence of 2.9% among ART-naive chronic HIV-1 infected individuals.

Evaluation of an opa gene-based nucleic acid amplification test for detection of Neisseria gonorrhoeae in urogenital samples in North India.

Due to the poor positive predictive value of nucleic acid amplification tests (NAATs) for gonorrhoea when applied to a low-prevalence setting, current guidelines recommend the use of supplementary polymerase chain reaction (PCR) targeting a different gene for confirmation of true positives in urogenital specimens. This study sought to standardize and evaluate performance of an in-house opa gene-based PCR assay for gonorrhoea compared to assays targeting the porA pseudogene and 16S rRNA gene. Four hundred samples (300 endocervical, 100 urethral swabs) from patients attending STD clinics in New Delhi, India were used. The sensitivity, specificity, positive predictive value and negative predictive value of the opa-based PCR were 100%, 97·9%, 89·5% and 100%, respectively. In females, the use of NAATs provided enhanced diagnosis of gonorrhoea.

Pneumocystis jirovecii dihydropteroate synthase (DHPS) genotypes in non-HIV-immunocompromised patients: a tertiary care reference health centre study.

Studies on Pneumocystis jirovecii dihydropteroate synthase (DHPS) genotypes among non-HIV immunocompromised patients from developing countries are rare. In the present prospective investigation, 24 (11.8%) cases were found to be positive for Pneumocystis jirovecii out of 203 non-HIV patients with a clinical suspicion of Pneumocystis pneumonia (PCP). Dihydropteroate synthase (DHPS) genotype 1 (Thr55+Pro57) was noted in 95.8% P. jirovecii isolates in the present study in contrast to only 4.1% of patients with DHPS genotype 4 (Thr55Ala + Pro57Ser).

Acanthamoeba keratitis with Curvularia co-infection.

We report a case of Acanthamoeba keratitis with Curvularia co-infection. Acanthamoeba and fungal co-infection have been uncommonly reported in literature, worldwide. A classical history with a strong clinical suspicion and experienced laboratory personnel with systematic examination of corneal scrapings for bacterial, viral, parasitic and fungal causes are imperative for accurate diagnosis. Early diagnosis of Acanthamoeba keratitis or fungal infection followed by aggressive and appropriate treatment with effective agents is critical for the retention of good vision. Acanthamoeba keratitis is difficult to diagnose and, despite improvement in treatment options, may culminate in prolonged morbidity and significant loss of visual acuity. This case emphasizes the important role played by clinical microbiologists in making prompt diagnosis which can ultimately reduce visual morbidity.

Dirofilariasis: a rare case report.

Human dirofilariasis is a zoonotic infection most commonly caused by Dirofilaria repens. It has not been widely recognized in India. There is probably a focus of human infection with D. repens in Kerala. We report the first case of dirofilariasis, from the Eastern-part of India, to the best of our knowledge. Among the documented cases of human dirofilariasis caused by D. repens, recorded in India, most of them had ocular infections and few had subcutaneous involvement of the face. This is the first case report of human dirofilariasis from India involving the lower part of human body.

Clinical and molecular characteristics of nosocomial meticillin-resistant Staphylococcus aureus skin and soft tissue isolates from three Indian hospitals.

We analysed risk factors for nosocomial meticillin-resistant Staphylococcus aureus (MRSA) skin and soft tissue infections (SSTIs) in three Indian hospitals. We also determined antimicrobial resistance patterns and genotypic characteristics of MRSA isolates using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and staphylococcal cassette chromosome (SCCmec) typing. Medical records of 709 patients admitted to three tertiary hospitals with nosocomial S. aureus SSTIs were clinically evaluated. Antimicrobial susceptibility testing of patient isolates was performed in accordance with Clinical and Laboratory Standards Institute guidelines, with meticillin and mupirocin resistance confirmed by multiplex polymerase chain reaction. PFGE analysis of 220 MRSA isolates was performed, followed by MLST and SCCmec typing of a selected number of isolates. MRSA was associated with 41%, 31% and 7.5% of infections at the three hospitals, respectively. Multiple logistic regression analysis identified longer duration of hospitalisation [odds ratio (OR): 1.78; OR: 2.83 for >or=20 days], intra-hospital transfer (OR: 1.91), non-infectious skin conditions (3.64), osteomyelitis (2.9), neurological disorders (2.22), aminoglycoside therapy (1.74) and clindamycin therapy (4.73) as independent predictors for MRSA SSTIs. MRSA isolates from all three hospitals were multidrug resistant, with fifteen clones (I-XV) recognised. A majority of the strains possessed type III cassette. The common sequence type (ST) 239 was considered the signature MLST sequence for PFGE clone III. This major MRSA clone III was closely related to the UK EMRSA-1 and was significantly more resistant to antibiotics. Dissemination of multidrug-resistant MRSA clones warrants continuous tracking of resistant genotypes in the Indian subcontinent.

Outbreak of meningococcal disease in and around New Delhi, India, 2005-2006: a report from a tertiary care hospital.

The first of several cases of meningococcal meningitis was reported in April 2005, in New Delhi, India. Subsequent to this the Government declared an outbreak, which persisted for two periods, from April-July 2005 and January-March 2006. The National Institute of Communicable Diseases (NICD) recommended using WHO criteria for diagnosis of disease. During the outbreak 380 clinically suspected cases were investigated. Of 55 cases diagnosed as confirmed/probable the mortality rate was 14.6%. Meningitis was reported in 60% of cases and meningococcaemia in 40%. Microscopy of petechial rash was positive in 87.5%, CSF Gram stain positive in 68.3%, and latex agglutination test of CSF positive in 64.9% of samples. Neisseria meningitidis (serogroup A) was isolated from 37.7% of cases, 57.7% from CSF. Blood culture was positive in 10.4% of cases. CrgA polymerase chain reaction for N. meningitidis confirmed the isolates. All isolates were susceptible to third-generation cephalosporins, azithromycin and rifampicin, with increasing resistance to ceftriaxone. Penicillin resistance was encountered in 15.4% of strains. Resistance to quinolones was very high at 100% for levofloxacin, 84.6% for ofloxacin and 65.4% for ciprofloxacin. All patients with penicillin-resistant organisms (4) or intermediate sensitivity (4) succumbed to the disease. These patients also had a higher minimum inhibitory concentration to ceftriaxone.

Parasites in patients with malabsorption syndrome: a clinical study in children and adults.

BACKGROUND: Intestinal parasites not only cause diarrheal diseases but also significant malabsorption. Literature on the role of parasites, such as intestinal coccidia and microsporidia in malabsorption syndrome is limited. METHODS: Three consecutive stool samples from 50 adult and 50 children patients with malabsorption syndrome and an equal number of healthy controls without diarrhea were examined for intestinal coccidia, microsporidia and other intestinal parasites by wet mount, Kinyoun's modified acid-fast staining and chromotrope 2R staining. RESULTS: Celiac disease was the commonest cause of malabsorption syndrome in both adults (52%) and children (74%). Forty (80%) and 41 (82%) adults and children, respectively, with malabsorption syndrome were infected with parasites. These results were significantly higher in comparison to those from the healthy adults and children controls (22% and 16%), respectively (P < 0.001). Of them, 48% and 46% of the adults and children, respectively, with malabsorption had pathogenic parasitic infections. The pathogenic parasites detected in adults were Giardia lamblia 12 (24%), E. histolytica / dispar 5 (10%), Ancylostoma duodenale 4 (8%), H. nana 2 (4%) and Cyclospora cayetanensis 1 (2%). The pathogenic parasites detected in children with malabsorption syndrome were Giardia lamblia 8 (16%), Cryptosporidium 7 (14%), E. histolytica / dispar 3 (6%), Ancylostoma duodenale 3 (6%), Isospora belli 1 (2%), and H. nana 1 (2%). None of the stool samples from healthy controls were positive for Cryptosporidium spp., Cyclospora and Isospora belli. All the patients infected with intestinal coccidia were HIV sero-negative. CONCLUSION: Celiac disease is the most common cause of malabsorption syndrome in both adults and children. These people harbor significantly more pathogenic parasites and are more frequently colonized with harmless commensals as compared to healthy controls. Intestinal coccidia are associated with malabsorption syndrome, particularly in malnourished children.

Fatal haemophagocytic syndrome and hepatitis associated with visceral leishmaniasis.

Haemophagocytic syndrome (HPS) secondary to infections occurs due to excessive, non-malignant proliferation of histiocytes, with resultant haemophagocytosis. The syndrome is essentially treatable, provided timely etiological diagnosis is achieved. In this report, we present a rare case of a child who hailed from Uttaranchal and presented with severe hepatitis. Bone marrow examination revealed an unexpected diagnosis of HPS secondary to visceral leishmaniasis. Despite initiating appropriate antileishmanial treatment, the child had a fatal outcome.