Seroprevalence of transplacentally acquired measles antibodies in HIV-exposed versus HIV-unexposed infants at six months of age.

BACKGROUND & OBJECTIVES: Measles infection is reported to be more severe, prolonged and associated with a higher complication rate in children with HIV infection. Reports indicate that infants born to HIV-infected women [HIV exposed infants (HEI)] may be more vulnerable to measles. The World Health Organization recommends measles vaccination starting at six months of age in these infants who may be HIV-infected themselves. However, in India, they are given measles vaccination at nine months of age like all other infants. In this study, the seroprevalence of transplacentally acquired measles antibodies was compared in HEI and unexposed infants (HUnI) at six months of age and the proportion of HEI undergoing seroconversion after immunization with measles vaccine was assessed. METHODS: In this prospective longitudinal study, measles IgG antibodies were estimated in serum of 49 HEI and 50 HUnI aged 6-7 months. Measles vaccine was then administered to HEI. Assessment for measles IgG antibodies was repeated 8-12 wk post-immunization. RESULTS: Measles IgG antibodies were detected in two of 49 (4.1%) HEI and 16 of 50 (32%) HUnI. HEI were 11 times more likely to lack measles antibodies as compared to HUnI (odds ratio=11.05, 95% confidence interval=2.989-40.908). Post-vaccination, seroprevalence of measles antibodies increased to 38.5 per cent (PInterpretation & conclusions: Most HEI lacked measles antibodies at six months age and were, therefore, more vulnerable to measles than HUnI. Seroconversion in response to a single dose of measles vaccine administered at six months age was low in these infants, signifying the need of additional dose(s) of measles/measles-containing vaccine.

New Delhi metallo-ß-lactamase - type carbapenemases producing Escherichia coli isolates from hospitalized patients: A pilot study.

BACKGROUND & OBJECTIVES: Resistances to carbapenem group of antimicrobials among Escherichia coli due to production of carbapenemases, especially the New Delhi metallo-ß-lactamase (NDM) types, pose serious challenges in the treatment of infections in healthcare settings. This study was undertaken to detect NDM producing E. coli isolates from hospitalized patients with urinary tract infection (UTI). METHODS: A total of 30 non-repetitive isolates of E. coli from hospitalized patients with clinical suspicion of UTI were subjected to antimicrobial susceptibility testing. Screening for the production of extended-spectrum ß-lactamases (ESBL) was carried out by minimum inhibitory concentration (MIC) test strip ESBL followed by phenotypic confirmation by double-disc synergy test. Phenotypic confirmation of carbapenemase production was carried out by MIC test strip metallo-ß-lactamases. Molecular identification of the blaNDM gene was carried out by polymerase chain reaction (PCR) and sequencing of the amplified fragment. RESULTS: Seventeen of the 30 isolates were detected as ESBL producers, of which three were found to be carbapenemase producers. NDM genes were detected by PCR followed by gene sequencing in all three isolates positive for ESBL as well as carbapenemase. The amino acid sequence of the three isolates showed complete identity to the reference sequences of NDM-1, NDM-4 and NDM-8, respectively. INTERPRETATION & CONCLUSIONS: Our study showed the circulation of NDM variants among the clinical isolates of E. coli that were producers of ESBL as well as carbapenemase.

Incidence, Etiology and Risk Factors Associated with Neonatal Healthcare-Associated Conjunctivitis: A Prospective Study from a Tertiary Care Hospital in India.

INTRODUCTION: Healthcare-associated conjunctivitis (HAC) can lead to serious sequelae including blindness. We conducted a one-year prospective study to determine the epidemiology of neonatal HAC at a tertiary-care hospital in India. METHODS: From the neonates fulfilling a set of predefined inclusion criteria, cases of HAC were diagnosed based on CDC guidelines. Conjunctival swabs, obtained from neonates with suggestive clinical signs, were processed using standard protocols. Twenty-eight potential risk factors were analyzed. RESULTS: We detected 24 cases of HAC among 591 enrolled neonates, with Escherichia coli being the most frequently isolated microorganism. On multivariate analysis, intubation at birth (p = 0.046) and orogastric feeding (p = 0.029) had a statistically significant association with neonatal HAC. Average hospitalization increased from 9.6 to 20.8 days for neonates diagnosed with HAC. CONCLUSION: A standardized case-definition and physician awareness of potential serious sequelae would help improve detection rates and timely institution of therapy. Hand hygiene could help control the menace of neonatal HAC.

Outbreaks of unexplained neurologic illness - Muzaffarpur, India, 2013-2014.

Outbreaks of an unexplained acute neurologic illness affecting young children and associated with high case-fatality rates have been reported in the Muzaffarpur district of Bihar state in India since 1995. The outbreaks generally peak in June and decline weeks later with the onset of monsoon rains. There have been multiple epidemiologic and laboratory investigations of this syndrome, leading to a wide spectrum of proposed causes for the illness, including infectious encephalitis and exposure to pesticides. An association between illness and litchi fruit has been postulated because Muzaffarpur is a litchi fruit-producing region. To better characterize clinical and epidemiologic features of the illness that might suggest its cause and how it can be prevented, the Indian National Centre for Disease Control (NCDC) and CDC investigated outbreaks in 2013 and 2014. Clinical and laboratory findings in 2013 suggested a noninflammatory encephalopathy, possibly caused by a toxin. A common laboratory finding was low blood glucose (<70 mg/dL) on admission, a finding associated with a poorer outcome; 44% of all cases were fatal. An ongoing 2014 investigation has found no evidence of any infectious etiology and supports the possibility that exposure to a toxin might be the cause. The outbreak period coincides with the month-long litchi harvesting season in Muzaffarpur. Although a specific etiology has not yet been determined, the 2014 investigation has identified the illness as a hypoglycemic encephalopathy and confirmed the importance of ongoing laboratory evaluation of environmental toxins to identify a potential causative agent, including markers for methylenecyclopropylglycine (MCPG), a compound found in litchi seeds known to cause hypoglycemia in animal studies. Current public health recommendations are focused on reducing mortality by urging affected families to seek prompt medical care, and ensuring rapid assessment and correction of hypoglycemia in ill children.

Chromoblastomycosis Caused by Cladophialophora bantiana in a Renal Transplant Recipient From Delhi, India.

A 45-year-old accountant residing in Delhi, India, presented to our dermatology clinic with a small asymptomatic plaque on the little finger of his left hand of 3 months' duration. The onset of the lesion was insidious and gradually progressed to 4 cm across at the time of his first visit. The patient had undergone renal transplantation twice (the first procedure 3 months prior and the second 18 months prior). Since then, he had been receiving cyclosporine A (400 mg daily) and prednisolone (40 mg) daily in immunosuppessive doses. The patient denied any kind of cutaneous injury prior to the onset of the lesion and any similar lesions in the past.

Gold-mercaptopropionic acid-polyethylenimine composite based DNA sensor for early detection of rheumatic heart disease.

The first gold-mercaptopropionic acid-polyethylenimine composite based electrochemical DNA biosensor was fabricated for the early detection of Streptococcus pyogenes infection in humans causing rheumatic heart disease (heart valve damage). No biosensor is available for the detection of rheumatic heart disease (RHD). Therefore, the mga gene based sensor was developed by the covalent immobilization of a 5'-carboxyl modified single stranded DNA probe onto the gold composite electrode. The immobilized probe was hybridized with the genomic DNA (G-DNA) of S. pyogenes from throat swabs and the electrochemical response was measured by cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance (EI). Covalent immobilization of the probe onto the gold composite and its hybridization with G-DNA was characterized by FTIR and SEM. The sensitivity of the sensor was 110.25 µA cm(-2) ng(-1) with DPV and the lower limit of detection was 10 pg per 6 µL. The sensor was validated with patient throat swab samples and results were compared with available methods. The sensor is highly specific to S. pyogenes and can prevent damage to heart valves by the early detection of the infection in only 30 min.

nspA gene as a specific genetic marker for detection of Neisseria meningitidis causing bacterial meningitis.

Bacterial meningitis caused by Neisseria meningitidis which causes human brain meninges damage, is generally diagnosed from patient cerebrospinal fluid through microscopy, immunological assays, biochemical test, PCR, microarray and biosensors. However, these methods are expensive, time-consuming or non-confirmatory due to certain limitations. A quick PCR based method was developed for detection of bacterial meningitis caused by N. meningitidis using specific primers based on amplification of virulence nspA (Neisseria surface protein A) gene partial sequence (202 bp). The nspA gene amplicon could be used as a genetic marker for minimum detection of 10 ng genomic DNA (G-DNA) of N. meningitidis with high sensitivity only in 80 min, which is least time reported for the confirmation of the disease. However, the lower detection limit was found as low as 1.0 ng G-DNA, but with less sensitivity. The cross-reactivity of the genetic marker, was also studied with other possible pathogens. A comparison with the presently available detection methods and our method was also done using patient samples.

Carbon-Mercaptooctadecane/Carboxylated Multi-walled Carbon Nanotubes Composite Based Genosensor for Detection of Bacterial Meningitis.

Human brain bacterial meningitis is a life-threatening disease mainly caused by Neisseria meningitidis, lead to several complications including damage of brain or even death. The present available methods for diagnosis of meningitis have one or more limitations. A rmpM gene based genosensor was fabricated by immobilizing 5'-amino modified 19-mer single stranded DNA probe onto carbon-mercaptooctadecane/carboxylated multi-walled carbon nanotubes composite electrode and hybridized with 2.5-40 ng/6 µL of single stranded genomic DNA (ssG-DNA) of N. meningitidis from cerebrospinal fluid (CSF) of the suspected meningitis patients. The electrochemical response was measured by using cyclic voltammetry and differential pulse voltammetry (DPV) using 1 mM methylene blue as redox indicator in 30 min (including a response time of 1 min) at 25 °C. The sensitivity of the genosensor was 3.762 (µA/cm(2))/ng and limit of detection was 2 ng of ssG-DNA of N. meningitidis with DPV. The genosensor has specificity only to N. meningitidis and does not hybridize with the genomic DNA of any other possible pathogen in human CSF. The immobilization of the probe and hybridization of the ssG-DNA were characterized by using electrochemical impedance in presence of 5 mM potassium ferricyanide and scanning electron microscopy. The genosensor loses only 12 % of its original DPV current on storage at 4 °C for 6 months. Carbon composite based electrochemical array can be constructed to detect multiple bacterial meningitis suspected patient CSF samples during an outbreak of the disease.

Omp85 genosensor for detection of human brain bacterial meningitis.

The 5'-thiolated DNA probe based on specific virulence gene, Omp85, was immobilized onto a screen-printed gold electrode followed by hybridization with 6-100 ng/6 µl (5.9 × 10(5)-9.3 × 10(6 )c.f.u.) of Neisseria meningitidis single stranded genomic DNA (ssG-DNA) for 10 min at 25 °C from the cerebrospinal fluid (CSF) of a meningitis patient. The Omp85 genosensor can detect as little as 6 ng ssG-DNA in 6 µl CSF of a human brain meningitis patient in 30 min including a response time of 1 min by cyclic voltammetry, differential pulse voltammetry (DPV) and electrochemical impedance. The sensitivity of the genosensor electrode was 2.6(µA/cm(2))/ng using DPV with regression coefficient (R(2)) 0.954. The genosensor was characterized using Fourier transform infrared spectroscopy and atomic force microscopy. Omp85 genosensor was stable for 12 months at 4 °C with 12 % loss in DPV current.

Quick diagnosis of human brain meningitis using omp85 gene amplicon as a genetic marker.

The usual diagnosis of life-threatening human brain bacterial meningitis are expensive, time consuming or non-confirmatory. A quick PCR based diagnosis of meningitis in cerebrospinal fluids (CSF) using specific primers of virulent Omp85 gene of Neisseria meningitidis can detect as low as 1.0 ng of genomic DNA (G-DNA) in 80 min for confirmation of bacterial meningitis caused by N. meningitidis infection. The 257 bp amplicon of Omp85 gene does not show homology with other suspected pathogens in CSF and can be used as a specific genetic marker for diagnosis of the disease.

Correlation of partial env gene sequences with disease progression parameters in HIV-positive pregnant women from India.

Ever since the beginning of the epidemic of HIV, one of the poignant aspects of HIV infection is transmission of the virus from mother to child. It is not known whether pregnancy accelerates the progression of HIV infection from a clinically asymptomatic stage to a progressive clinical phase. Present study was carried out to understand disease progression in pregnant women from India. We studied co-receptor utilization (the major determinant of HIV disease progression), N-glycosylation sites, and sequence variability. Blood samples were collected from 25 HIV sero-positive patients, eleven from the antenatal risk group (experimental group), nine from heterosexual male, and five from heterosexual female risk group (control group). Partial env gene was amplified by PCR and sequenced. BLAST search and phylogenetic analysis were used to determine the subtype. The deduced amino acid sequence of the V3 region was used to predict co-receptor, determine sequence variability and N-glycosylation site. The experimental group comprising the antenatal risk group did not exhibit any difference in terms of co-receptor, N-glycosylation, and sequence variability when compared with the control, non-pregnant group. Pregnancy does not seem to accelerate the clinical course of HIV infection. The female body during the gestation phase possibly acquires certain strategies to impede or at least alleviate the disease progression during the crucial immune-compromised pregnancy phase, which would otherwise adversely affect the mother as well as the fetus during the infection.

Emergence of pandemic 2009 influenza A H1N1, India.

BACKGROUND & OBJECTIVES: Pandemic H1N1 caused deluge of cases from 74 countries and prompted World Health Organization to raise warning to phase 6. The present study was conducted on throat and nasal swab samples received and tested at National Centre for Disease Control, Delhi, India during 2009-2010 to collect epidemiological and clinical information on positive cases. METHODS: Throat and nasopharyngeal swabs from category C influenza A H1N1 patients during May 2009-September 2010 along with their clinico-epidemiological details were collected from identified hospitals from Delhi and other States. Samples were tested by Real time reverse transcriptase PCR using primers and probes developed at CDC, Atlanta for four influenza target genes. RESULTS: A total of 33,751 samples, both throat and nasal swab samples from each patient were tested for H1N1 influenza virus, of which, 7943 (23.5%) were positive for pandemic influenza A H1N1 and 3759 (11.1%) were positive for influenza A (seasonal flu). Maximum number of positive cases (N=2792, 35.1%) were from 20-39 yr age group, comprising 1790 (22.5%) males and 1182 (14.8%) females. Only 2620 (33%) positive cases were close contact of influenza A H1N1 positive patient. Majority cases presented (N=2792, 35.1%) with fever 7005 (88.1%), followed by 6133 cases (77.2%) exhibiting fever and cough, 377 (4.7%) complained of fever, cough, nasal catarrh and 362 (4.5%) cases had fever with shortness of breath. INTERPRETATION & CONCLUSIONS: The study showed a peak of cases of pandemic influenza A H1N1 in December 2009 and indicated predominance of H1N1 positive cases among 20-39 yr age group and among males compared to females.

Whole cell pertussis vaccine: reducing toxicity.

Since introduction of the pertussis vaccine in 1940's the morbidity and mortality due to the infection has been markedly reduced all over the world. However the adverse effects of the inactivated whole cell pertussis vaccine like pain, swelling at the site of injection, fever, vomiting anorexia, persistent crying & drowsiness have been the cause of great concern, till date. Also the safety concerns over the use of thiomersal as an inactivating agent as well preservative have been raised in the recent past. Studies in many countries have been initiated to reduce or replace thiomersal & using other inactivating agents in the vaccines. Limited studies have been conducted in India. The present study has been undertaken as an attempt to reduce the quantity of thiomersal and modification in the procedure of production of the pertussis vaccine to reduce the toxicity, to produce better quality of whole cell pertussis vaccine. To achieve this, at the time of production of the whole cell pertussis vaccine, at Kasauli, as per the standard procedure recommended by WHO, three parallel batches of the pertussis culture were inactivated in fermenter before harvesting and thiomersal was used only one time for suspending the bacterial mass after harvesting. The resultant modified vaccine so prepared when tested showed that it was of better quality as compared to the one produced by standard procedure, when tested in mice.

Quick PCR based diagnosis of typhoid using specific genetic markers.

A quick multiplex PCR based detection method was developed for early diagnosis of typhoid using specific genetic markers of S. typhi. Primers of tyv gene, flag gene, viaB gene and ratA gene confirmed the specificity and sensitivity of the PCR. The serum samples of the suspected typhoid patients were taken directly for PCR without culturing and isolating genomic DNA. Overall diagnosis required 2 h which is the least time ever reported for a PCR based methods. The sensitivity of the method is up to 5 fg genomic DNA. The genetic markers are specific and the four pairs of primers give selective amplification and differentiate S. typhi from closely related S. typhimurium.

Impact of HIV on genomic variability in the 5'UTR of HCV in Indian patients with HCV/HIV co-infection.

BACKGROUND: The impact of HIV on hepatitis C virus (HCV) genome during HCV/HIV co-infection is poorly understood. The present study was intended to unveil nucleotide sequence variability in the 5'-untranslated region (5'UTR) of HCV in co-infected cases. METHODS: Automated nucleotide sequencing of the 5'UTR of HCV from both mono- and co-infected cases was performed. RESULTS: Data analysis revealed deletion of a continuous stretch of 12 nucleotides (nt 240-251) from domain IIIc in 20% co-infected cases, but no long-stretch deletion was observed in HCV from mono-infected cases. On the contrary, there was no insertion in the 5'UTR of HCV from co-infectedcases, but there were insertions in domain II and III (3 mononucleotides and 2 dinucleotides) of the 5'UTR in mono-infected cases. CONCLUSION: Since domain III is known to be important for binding of 40S ribosomal subunit, deletion of a single stretch of 12 nucleotides in HCV from co-infected cases observed in the present study may have implications during HCV replication with or without HIV infection. Although this is the first report on genomic heterogeneity in the 5'UTR of HCV from HCV/HIV co-infected Indian patients, it would be worthwhile to study if similar changes are observed in other genes of HCV during co-infection.

Epidemiological Profile of Acute Viral Encephalitis.

OBJECTIVE: To study the etiology and clinico-epidemiological profile of acute viral encephalitis in children with acute encephalitis syndrome (AES). METHODS: An observational study including 100 patients fulfilling the criteria for AES was conducted in children of age group 1 mo - 16 y. Viral isolation was done on RD cells, HEp-2 cells and Vero cells from the cerebrospinal fluid samples of suspected viral encephalitis (VE) cases. An enzyme immunoassay for IgM antibodies was performed for measles, mumps, Varicella zoster virus (VZV), Herpes simplex virus 1 (HSV1) and Japanese encephalitis virus (JEV). Multiplex polymerase chain reaction (PCR) was done for Cytomegalovirus, Epstein Barr virus (EBV), HSV1 & 2, VZV, Enterovirus, Parecho virus, Human Herpes virus (HHV 6, 7) and Parvovirus B19. A micro neutralization test was performed for Enterovirus 71. RESULTS: Out of enrolled 100 patients, 73 were of probable viral encephalitis. HSV1 (31.50%) was the commonest virus followed by Adenovirus (10.95%), Parvovirus (2.73%), JE virus (1.36%), Enterovirus (1.36%), EBV (1.36%), and mixed infection with HSV & EBV (1.36%). HSV 1 caused significant morbidity in children. The common computed tomography (CT) findings were hypodensities in the fronto- parietal lobe followed by cerebral edema. CONCLUSIONS: The landscape of AES in India has changed in the previous decade, and both outbreak investigations and surveillance studies have increasingly reported non-JEV etiologies; because of these findings there is a need to explore additional strategies to prevent AES beyond vector control and JEV vaccination.

Outbreak of Coxsackie B4 arthritis among newborns and staff of a neonatal unit.

We investigated an outbreak of Coxsackie B4 arthritis in a neonatal unit involving 20 neonates and 12 staff members, over an eight-month period. Laboratory investigations, serology tests, indicate that the outbreak was caused by Coxsackie B4 virus. Contamination of one of the overhead water reservoirs, supplying the nursery, was responsible. After the water tanks were cleaned out, no new cases were reported over five years.

DNA chip based sensor for amperometric detection of infectious pathogens.

Several infectious pathogens are found in human whose detection is essential for rapid cure of diseases. The most commonly found pathogen in human is Streptococcus pyogenes which leads to a wide range of infections from mild pharyngitis to rheumatic heart disease. An ultrasensitive DNA chip based sensor was developed for quick identification of pathogen S. pyogenes from patient throat swab samples. The amperometric response was measured after hybridization of specific probe with single stranded genomic DNA (ssG-DNA) from the patient samples. The DNA chip was characterized by FTIR, SEM and validated with suspected patient real samples. The sensitivity of the DNA chip based sensor was found 951.34(µA/cm2)/ng DNA and lower limit of detection (LOD) was 130fg/6µL samples. The DNA chip based sensor is highly specific and takes only 30min for identification of specific pathogen.

Association of acute toxic encephalopathy with litchi consumption in an outbreak in Muzaffarpur, India, 2014: a case-control study.

BACKGROUND: Outbreaks of unexplained illness frequently remain under-investigated. In India, outbreaks of an acute neurological illness with high mortality among children occur annually in Muzaffarpur, the country's largest litchi cultivation region. In 2014, we aimed to investigate the cause and risk factors for this illness. METHODS: In this hospital-based surveillance and nested age-matched case-control study, we did laboratory investigations to assess potential infectious and non-infectious causes of this acute neurological illness. Cases were children aged 15 years or younger who were admitted to two hospitals in Muzaffarpur with new-onset seizures or altered sensorium. Age-matched controls were residents of Muzaffarpur who were admitted to the same two hospitals for a non-neurologic illness within seven days of the date of admission of the case. Clinical specimens (blood, cerebrospinal fluid, and urine) and environmental specimens (litchis) were tested for evidence of infectious pathogens, pesticides, toxic metals, and other non-infectious causes, including presence of hypoglycin A or methylenecyclopropylglycine (MCPG), naturally-occurring fruit-based toxins that cause hypoglycaemia and metabolic derangement. Matched and unmatched (controlling for age) bivariate analyses were done and risk factors for illness were expressed as matched odds ratios and odds ratios (unmatched analyses). FINDINGS: Between May 26, and July 17, 2014, 390 patients meeting the case definition were admitted to the two referral hospitals in Muzaffarpur, of whom 122 (31%) died. On admission, 204 (62%) of 327 had blood glucose concentration of 70 mg/dL or less. 104 cases were compared with 104 age-matched hospital controls. Litchi consumption (matched odds ratio [mOR] 9·6 [95% CI 3·6 - 24]) and absence of an evening meal (2·2 [1·2-4·3]) in the 24 h preceding illness onset were associated with illness. The absence of an evening meal significantly modified the effect of eating litchis on illness (odds ratio [OR] 7·8 [95% CI 3·3-18·8], without evening meal; OR 3·6 [1·1-11·1] with an evening meal). Tests for infectious agents and pesticides were negative. Metabolites of hypoglycin A, MCPG, or both were detected in 48 [66%] of 73 urine specimens from case-patients and none from 15 controls; 72 (90%) of 80 case-patient specimens had abnormal plasma acylcarnitine profiles, consistent with severe disruption of fatty acid metabolism. In 36 litchi arils tested from Muzaffarpur, hypoglycin A concentrations ranged from 12·4 µg/g to 152·0 µg/g and MCPG ranged from 44·9 µg/g to 220·0 µg/g. INTERPRETATION: Our investigation suggests an outbreak of acute encephalopathy in Muzaffarpur associated with both hypoglycin A and MCPG toxicity. To prevent illness and reduce mortality in the region, we recommended minimising litchi consumption, ensuring receipt of an evening meal and implementing rapid glucose correction for suspected illness. A comprehensive investigative approach in Muzaffarpur led to timely public health recommendations, underscoring the importance of using systematic methods in other unexplained illness outbreaks. FUNDING: US Centers for Disease Control and Prevention.