Patients In Kolkata Research Articles

Incidence, Virulence Factors, and Clonality among Clinical Strains of Non-O1, Non-O139 Vibrio cholerae Isolates from Hospitalized Diarrheal Patients in Kolkata, India

The incidence of Vibrio cholerae non-O1, non-O139 strains from hospitalized patients with acute diarrhea constituted 27.4% (n = 54) of the total 197 V. cholerae strains isolated from patients in Kolkata, India, in 2003. Of 197 strains, 135 were identified as O1 serotype Ogawa and 2 were identified as O139. In the same time period, six O1 background rough strains that possessed all known virulence factors were identified. Serotype analysis of the non-O1, non-O139 strains placed 42 strains into 19 serogroups, while 12 remained O nontypeable (ONT); the existing serotyping scheme involved antisera to 206 serogroups. Detection of a good number of ONT strains suggested that additional serogroups have arisen that need to be added to the current serotyping scheme. The non-O1, non-O139 strains were nontoxigenic except for an O36 strain (SC124), which regulated expression of cholera toxin as O1 classical strains did. Additionally, strain SC124 carried alleles of tcpA and toxT that were different from those of the O1 counterpart, and these were also found in five clonally related strains belonging to different serogroups. Strains carrying tcpA exhibited higher colonization in an animal model compared to those lacking tcpA. PCR-based analyses revealed remarkable variations in the distribution of other virulence factors, including hlyA, rtxA, Vibrio seventh pandemic island I (VSP-I), VSP-II, and type III secretion system (TTSS). Most strains contained hlyA (87%) and rtxA (81.5%) and secreted cytotoxic factors when grown in vitro. Approximately one-third of the strains (31.5%) contained the TTSS gene cluster, and most of these strains were more motile and hemolytic against rabbit erythrocytes. Partial nucleotide sequence analysis of the TTSS-containing strains revealed silent nucleotide mutations within vcsN2 (type III secretion cytoplasmic ATPase), indicating functional conservation of the TTSS apparatus.

Immediate hypersensitivity to common food allergens-An investigation on respiratory allergic patients in Kolkata (Calcutta), India

Immediate hypersensitivity to common food allergens-An investigation on respiratory allergic patients in Kolkata (Calcutta), India

Molecular characterization of recent Vibrio cholerae O1, El Tor, Inaba strains isolated from hospitalized patients in Kolkata, India

To study the phenotypic and genotypic characterization of newly emerged V. cholerae O1, Inaba strains isolated from patients with diarrhoea. Bacterial characterization was made using polymerase chain reaction, ribotyping, PFGE and RFLP. After its first appearance in July 2004, O1 Inaba became the dominant serotype by March 2005 and totally replaced the former dominant serotype, Ogawa from May 2005. Most of the Inaba isolates belong to a new ribotype RIV. Ogawa and also some Inaba strains isolated during the same period were identified as RIII. Similarly, the majority of the Inaba isolates belong to 'H1' pulsotype and one isolate is type 'H', while the Ogawa isolates were mostly 'H' pulsotype. Presence of CTX prophage was detected in a single site of the chromosome with at least two RS elements. There has been a switch of dominant serotype from Ogawa to Inaba in the Kolkata region. This is not necessarily due to emergence of a new clone but does serve as an epidemiological marker. Further analysis at the molecular level will be required to define this trend and to monitor future spread to other regions.

Intestinal parasitism and Vibrio cholerae infection among diarrhoeal patients in Kolkata, India

In this study, we have analysed the epidemiological significance of the concurrent infections caused by Vibrio cholerae and intestinal parasites among different age groups of hospitalized diarrhoeal patients in Kolkata. A total of 3556 stool samples collected during 1996-2004 were screened for vibrios and parasites. The seasonality of V. cholerae and parasitic infections were studied in detail. The detection rates for Ascaris lumbricoides and Giardia lamblia infection were more than for the hookworm, Trichuris trichiura and Entamoeba histolytica. V. cholerae O1 was identified as the predominant serogroup among diarrhoeal patients. The highest rates for V. cholerae infection were in the 2-5 years age group and combined infection of V. cholerae and parasites was recorded among children aged between 2 and 10 years.

Multilocus sequence typing and genetic structure of Cryptosporidium hominis from children in Kolkata, India

Endemicity of cryptosporidiosis in India has been documented with little genetic characterization of the parasites. Fifty Cryptosporidium-positive specimens collected between 2001 and 2004 from pediatric patients in Kolkata, India were analyzed for parasite genetic structure using multilocus sequence typing (MLST). Genotype analyses showed the presence of Cryptosporidium hominis, Cryptosporidium meleagridis and Cryptosporidium felis in 49, 2 and 1 patients, respectively (two patients had mixed infections of C. hominis and C. meleagridis). To assess the extent of genetic heterogeneity of C. hominis, minisatellites, microsatellites and polymorphic markers in three different chromosomes were sequenced, including genes encoding the 60 kDa glycoprotein (GP60), a 47 kDa protein (CP47), a mucin-like protein (Mucin1), a serine repeat antigen (MSC6-7), and a 56 kDa trans-membrane protein (CP56) in chromosome 6, the 70 kDa heat shock protein (HSP70) in chromosome 2, and a T-rich gene fragment (Chrom3T) in chromosome 3. Population sub-structure of C. hominis based on multilocus gene sequences showed that there were 25 multilocus subtypes defined by combined sequence length and nucleotide polymorphism, which formed four distinct groups in this population. Significant intra- and inter-genic linkage disequilibria were observed with minimum recombination or expansion of limited subtypes, all indicative of a mostly clonal population structure. The results highlight the importance of high resolution MLST in studying Cryptosporidium population sub-structure especially when length polymorphism may be inadequate in identifying unique subtypes. The significance of the diverse MLST within C. hominis in relation to geographical and temporal factors and clinical manifestations of disease warrants further investigations.

Distribution and characterization of integrons in various serogroups of Vibrio cholerae strains isolated from diarrhoeal patients between 1992 and 2000 in Kolkata, India

A total of 133 clinical strains of Vibrio cholerae comprising 44 strains of O1, 45 strains of O139 and 44 strains of non-O1, non-O139 serogroups isolated from hospitalized patients in Kolkata, India, from 1992 to 2000 was examined for the presence of class 1, 2 and 4 integrons. By PCR and DNA sequencing, seven strains of O1, one strain of O139 and six strains of non-O1, non-O139 serogroups were found to contain class 1 integron-harbouring genes aadA1, aadA2 (encoding resistance to streptomycin and spectinomycin), blaP1 (resistance to beta-lactams), aar-3 (resistance to rifampicin), aacA4 (resistance to kanamycin and gentamicin), and dfrA1 and dfrA15 (resistance to trimethoprim). Most strains produced one or two bands using primers specific for the amplification of the variable region where the antibiotic-resistance genes are located, and their sizes ranged from 700 to 1250 bp. However, one strain of V. cholerae O1 isolated in 1994 gave a 2483 bp fragment, the largest fragment so far found in a class 1 integron of V. cholerae O1. No strain was positive for the intI2 gene. All V. cholerae strains, regardless of serogroup, were positive for the intI4 gene by PCR and using a colony hybridization test. Amplification of the intI4 gene by PCR yielded a 2200 bp fragment (1260 bp larger than the expected size) from three V. cholerae O139 strains isolated in 1999. Sequence analysis of this amplicon revealed an insertion of IS1359 in the middle of the intI4 gene. These data indicate that a class 1 integron is present in some clinical strains of V. cholerae isolated in Kolkata, India, and that a class 4 integron is ubiquitously distributed among V. cholerae strains regardless of serogroup.

Molecular epidemiology of HCV infection among acute and chronic liver disease patients in Kolkata, India

In recent years, hepatitis C virus (HCV) infection is gaining importance in Asian countries. Recent studies conducted in different parts of the world revealed that there is a genotypic correlation of disease severity and treatment outcome. A detailed study was carried out to delineate the genotypic distribution of HCV among acute and chronic liver disease patients in Kolkata, a city in eastern India. Acute and chronic liver disease was diagnosed among patients attending hepatitis clinics in the city. Anti-HCV ELISA was performed on the blood samples of the cases and positive samples were tested for presence of HCV-RNA and genotyping of the samples were carried out by reverse transcription and polymerase chain reaction (RT-PCR) and sequencing. Seroprevalence of HCV infection among acute (11.0%) and chronic (25.3%) hepatitis patients were high and among them 97 (75.8%) and 323 (86.1%) were HCV-RNA positive for acute and chronic hepatitis patients, respectively. Genotyping by PCR showed that the predominant genotype was 3b (42.3%) followed by 3a (28.9%) among acute hepatitis group whereas among chronic hepatitis group, the most prevalent genotypes were 3a (34.7%) and 3b (47.7%). Sequence analysis of the untypeable isolates revealed the presence of a rare subtype 6b. The study revealed very high prevalence of HCV among acute and chronic hepatitis patients with predominance of genotype 3. Subtype 6b was commonly found in Thailand but not in India. The detection of this rare strain of Thai origin reveals the spread of HCV infection from Thailand to other parts of Asia. This observation necessitates further intensive surveillance of HCV infection in India to unravel the distribution of genotypes in the country and to correlate disease severity and treatment outcome to the genotype prevalence.