Browsing by Author "Sridharan, G"
Now showing 1 - 20 of 67
Results Per Page
Sort Options
Item Aciclovir resistance among Indian strains of Herpes simplex virus as determined using a dye uptake assay.(2007-07-29) Abraham, A M; Kavitha, S; Joseph, P; George, R; Pillay, D; Malathi, J; Jesudason, M V; Sridharan, GResistance to aciclovir (ACV) among Herpes simplex virus (HSV) isolates is increasingly being reported in the literature particularly in immunocompromised patients. However, there is only limited data available from India despite widespread use of ACV in our hospital. A cross-sectional study was hence conducted to determine the aciclovir (ACV) susceptibility of HSV 1 and 2 isolates using a dye uptake (DU) assay. This study showed a 3.0% prevalence of ACV resistance among HSV-1 strains (2/66, median IC 50 0.098 microg/mL) while in HSV-2 strains, it was 7.8% (5/64, median IC 50 0.195 microg/mL). The IC 50 for the HSV-1 and HSV-2 strains resistant to ACV was greater than or equal to 6.25 microg/mL.Item Antibody to group A streptococcal carbohydrate in rheumatic fever and rheumatic heart disease.(1983-02-01) Koshi, G; Sridharan, G; Thangavelu, C PItem Antigen sharing of Salmonella typhi non-flagellar proteins with other salmonellae and some shigellae and Escherichia coli.(1991-04-01) Jesudason, M; Sridharan, G; John, T JCathodal moving protein components were identified in agarose gel electrophoresis of the Veronal buffer extract of a non-motile strain of S. typhi (8393, Colindale). Rabbit antiserum was raised against these cationic proteins; it had both agglutinating and precipitating activity. A total of 80 salmonella strains belonging to serogroups A, B, C1, C2, D, E1 and E2 including 26 S. typhi and 10 S. paratyphi A were tested against this antiserum in a slide agglutination test; all strains were agglutinated. Among 94 other bacterial strains tested, the antiserum agglutinated all 16 strains of Shigella flexneri, 2 of 5 Shigella sonnei, 5 of 34 E. coli and 1 of 8 Citrobacter species. These results show that there is antigenic sharing of the non-flagellar proteins of S. typhi with many other salmonellae as well as with some shigellae and E. coli.Item C substance--specific latex agglutination for early & rapid detection of Streptococcus pneumoniae in blood cultures.(1995-12-01) Jesudason, M V; Sridharan, G; Arulselvan, K; Joseph, A; Steinhoff, M C; John, T JA slide agglutination test was developed using latex particles coated with antiserum against the C substance, a common antigen for all serotypes of Streptococcus pneumoniae. This test was used for the rapid identification of pneumococci in blood culture broths which contained Gram positive cocci (GPC) in pairs or short chains on smear examination. Of 238 consecutive blood cultures with GPC tested, 72 were positive for Strep. pneumoniae by the latex test and conventional methods. The remaining 166 cultures were negative for both these, indicating a 100 per cent specificity and sensitivity for the test.Item A case of triple retroviral infection with HIV-1 (genotype C3), HIV-2 and HTLV-I.(2000-09-24) Ramalingam, S; Kannangai, R; Prakash, K J; Jesudason, M V; Sridharan, G; Kannangay, RItem Characterization of antibody response to human cytomegalovirus in Indian renal transplant patients.(2001-06-31) Finny, G J; Rao, M; Mach, M; Juneja, R; Thomas, P P; Jacob, C K; Manayani, D J; Abraham, P; Abraham, M; Sridharan, GBACKGROUND & OBJECTIVES: Cytomegalovirus (CMV) disease in seroendemic transplant populations is due to reactivation of the virus, or reinfection. In this context, the antibody response is likely to influence presentation, clinical severity and outcome of the disease, and may provide a diagnostic and prognostic marker. This study was carried out in Indian renal transplant patients and healthy adults to characterize the antibody response to cytomegalovirus. METHODS: Thirty three transplant recipients with CMV illness (symptomatology with IgM and/or nPCR positive status), 20 recipients who were asymptomatic in the 6 months of follow up after transplantation and 62 healthy controls were investigated for markers of CMV infection. These individuals were tested for IgG avidity and neutralizing antibody by ELISA techniques. RESULTS: All 53 transplant recipients were found to have an IgG avidity index of > 50 per cent. Antibody to a CMV envelope glycoprotein gB/AD-1 (putative neutralizing antibody) was expressed as S/N ratio and was > or = 5 in asymptomatic (65%) and symptomatic (27%) immunosuppressed renal transplant recipients. However, none of the 53 CMV IgG positive healthy controls were positive for neutralizing antibodies S/N ratio > or = 5 (S/N ratio = sample mean OD/mean OD of 3 negative controls in each run). We observed the simultaneous presence of CMV PCR signal in leukocytes and neutralizing antibody (S/N ratio > or = 5) in the plasma in 22 (41.5%) of the 53 renal transplant recipients. INTERPRETATION & CONCLUSIONS: In this study among the immunosuppressed transplant patients we observed an association between symptomatic disease and the relative absence of neutralizing antibodies. The neutralizing antibodies are less frequently demonstrable among controls; while appearance in a higher proportion of asymptomatic recipients especially in association with high IgG avidity (> 90%) is suggestive of its role in control of CMV disease despite reactivation as evidenced by DNAemia while on immunosuppressive therapy.Item Coagglutination for diagnosis of bacterial infection.(1989-05-01) Lalitha, M K; Sridharan, G; John, MItem Coagglutination method in the rapid diagnosis of acute bacterial meningitis.(1988-05-01) Mercy, J; Sridharan, G; Steinhoff, M C; Pereira, S M; Jadhav, M; Pulimood, B M; Mathai, D; Lalitha, M KItem Community prevalence of antibodies to human immunodeficiency virus in rural and urban Vellore, Tamil Nadu.(2005-01-20) Kang, Gagandeep; Samuel, Reuben; Vijayakumar, T S; Selvi, S; Sridharan, G; Brown, David; Wanke, ChristineBACKGROUND: Human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) are becoming increasingly common in India. Currently, antenatal prevalence is a surrogate marker for HIV prevalence in the community. The association between antenatal and community prevalence of HIV needs to be validated so that estimates can be verified or adjusted appropriately. METHODS: A probability proportional to size cluster survey was conducted in the Kaniyambadi block of Vellore district and in the urban wards of Vellore town to estimate the prevalence of antibodies to rubella from August 1999 to February 2000. All personal identifier data from the serum samples were removed to yield a collection for which only the age and sex were known. Estimation of antibodies to HIV in sera from individuals between 15 and 40 years of age, was carried out by one screening ELISA and the reactive sera were further subjected to a supplementary test. RESULTS: We tested 1512 serum samples from subjects residing in rural areas and 1358 samples from those residing in urban areas. The seropositivity among rural samples was 0.66% and among urban samples 1.4%. The prevalence was almost equal among men and women and the youngest infected individual was 15 years old. CONCLUSION: The prevalence of HIV during the period of study was similar to the national surveillance data for Tamil Nadu based on antenatal women. HIV prevalence differs in urban and rural Tamil Nadu, with urban areas having a higher burden of the disease.Item Comparison of a conventional polymerase chain reaction with real-time polymerase chain reaction for the detection of neurotropic viruses in cerebrospinal fluid samples.(2011-04) Ramamurthy, M; Alexander, M; Aaron, S; Kannangai, R; Ravi, V; Sridharan, G; Abraham, A MPurpose : To compare a conventional polymerase chain reaction (PCR) and real-time PCR for the detection of neurotropic DNA viruses. Materials and Methods : A total of 147 cerebrospinal fluid (CSF) samples was collected from patients attending a tertiary care hospital in South India for a period from 2005 to 2008. All these samples were tested using a conventional multiplex/uniplex PCR and a real-time multiplex/uniplex PCR. This technique was used to detect a large number of herpes viruses responsible for central nervous system infections, including HSV-1, HSV-2, VZV, CMV and EBV and the polyoma virus JCV. Results : Overall, in the entire set of samples, the real-time PCR yielded 88 (59.9%) positives and conventional PCR had six (4.1%) positives. Conclusion : Our results suggest that the real-time PCR assay was more sensitive compared with the conventional PCR. The advantage of real-time PCR is that it can be performed much faster than conventional PCR. Real-time PCR is less time-consuming, less labour-intensive and also reduces the chance of contamination as there is no post-amplification procedure. In the entire study population, the major viruses detected using real-time PCR were EBV (34%), HSV-2 (10.8%) and VZV (6.8%).Item Comparison of IgM capture ELISA with a commercial rapid immunochromatographic card test & IgM microwell ELISA for the detection of antibodies to dengue viruses.(2002-02-26) Sathish, N; Manayani, D J; Shankar, V; Abraham, Mary; Nithyanandam, G; Sridharan, GBACKGROUND & OBJECTIVES: There is a need for a reliable test for the early diagnosis of dengue fever (DF), which is now active in many parts of India especially in the post monsoon months. This study evaluated two commercial tests with an assay available from a national laboratory in India to obtain information and to make a comparison among the three tests as to which will be the most suited for the detection of IgM antibodies to dengue virus. METHODS: An IgM capture ELISA (National Institute of Virology, Pune, India) was compared with two commercial tests, the PanBio Rapid Immunochromatographic Card Test (Brisbane, Australia) and the PanBio Microwell IgM ELISA for the detection of IgM antibodies to dengue virus. We tested 154 samples from individuals with febrile illnesses having DF--like symptoms. RESULTS: The NIV IgM capture ELISA (MAC-ELISA) showed a high positivity rate (38.9%) as compared to the PanBio Rapid (22.7%) and the PanBio IgM ELISA (20.7%). The true prevalence of disease, sensitivity and specificity of the three tests were estimated using 2LC latent class models using expectation-maximization (EM) algorithm. The NIV MAC-ELISA showed a high sensitivity (96%) as compared to PanBio Rapid (73%) and PanBio IgM ELISA (72%). A subset of 68 samples (of the 154 tested) were analyzed by the NIV MAC-ELISA for IgM antibodies additionally to Japanese encephalitis (JE) and West Nile (WN) of which 31 samples showed positivity to either one, two or all three flaviviruses. Out of the 8 samples which were positive for dengue IgM alone by the NIV MAC-ELISA, only 2 (25%) each were picked up by the other 2 tests. While out of 7 samples positive for IgM to all three flaviviruses IgM by the NIV MAC-ELISA, 5 (71%) were picked up by the other 2 tests. Of the 5 that were picked up by the PanBio tests, 3 had the highest absorbance values to WN by the NIV MAC-ELISA, indicating cross reactivity by PanBio tests. INTERPRETATION & CONCLUSION: The MAC-ELISA though a 3 day procedure, would be a valuable screening test for the detection of IgM to dengue in routine diagnostic laboratories because of its high sensitivity and specificity rates. The test uses specific viral antigens to detect IgM antibodies not only to dengue but also to JE and West Nile as a result of which IgM antibodies to all the 3 commonly encountered flaviviruses can be detected in a single run. It also has the advantage in that depending on the strength of the antibody units obtained to a specific flaviviral antigen, presumptive diagnosis as to which particular arboviral infection has occurred can be made in conjunction with clinical presentation.Item Comparison of latex agglutination inhibition test & bioassays for rapid estimation of serum gentamicin levels.(1986-12-01) Ambro, J; Sridharan, G; Lalitha, M KItem Could the products of Indian medicinal plants be the next alternative for the treatment of infections.(2011-04) Nandagopal, B; Sankar, S; Ramamurthy, M; Sathish, S; Sridharan, GIndian medicinal plants are now recognized to have great potential for preparing clinically useful drugs that could even be used by allopathic physicians. Traditionally, practitioners of Indian medicine have used plant products in powder, syrup or lotion forms, without identification, quantification and dose regulation, unlike their allopathic counterparts. The present review explores the immense potential of the demonstrated effect of Indian medicinal plants on microbes, viruses and parasites. In the present context, with the available talent in the country like pharmaceutical chemists, microbiologists, biotechnologists and interested allopathic physicians, significant national effort towards identification of an "active principle" of Indian medicinal plants to treat human and animal infections should be a priority.Item Cytotoxicity of non O1, non O139 Vibrios isolated from fresh water bodies in Vellore, south India.(1999-11-19) Balaji, V; Sridharan, G; Jesudason, M VThe samples of plankton, soil sediment and water from a pond, a lake and a moat respectively in and around Vellore were studied for environmental vibrios. Vibrios were isolated from all these specimens after enrichment in alkaline peptone water and subculture on selective media. Non O1, non O139 Vibrio cholerae, Aeromonas spp. and Plesiomonas spp. were isolated. There were no isolates of V. cholerae serogroup O1 and O139. Representative strains of non O1 and non O139 V. cholerae from environmental sources were tested for toxin production in Chinese hamster ovary (CHO) and Vero cell monolayer in microtitre plates. Thirty-three (91.7%) of the 36 strains tested demonstrated cytopathic effect (CPE) in both cell lines indicating their toxigenicity. PCR done on representative strains of non O1 and non O139 V. cholerae showed that none of the strains were positive for ctx A, tcp A-E and tcp C genes. These results indicate that these non agglutinating environmental vibrios produced cytotoxins other than the cholera toxin.Item Cytotoxin testing of environmental Aeromonas spp. in Vero cell culture.(2004-05-29) Balaji, V; Jesudason, Mary V; Sridharan, GBACKGROUND & OBJECTIVES: Aeromonas spp. are known to cause a variety of infections in humans and this organism has been isolated from a variety of sources including environmental sources. The pathogenicity of the environmental isolates in and around Vellore has not been studied. This study was conducted to determine the cytotoxicity of the Aeromonas spp. isolated from water bodies, soil sediments, plankton and sewers in and around Vellore. METHODS: Aeromonas spp. isolated from environmental sources were identified by standard procedures. Representative isolates of Aeromonas spp. were tested for cell free cytotoxic factor in tissue culture system. Undiluted and diluted cell free filtrates of isolates and known toxigenic and non-toxigenic bacteria were added to Vero cell monolayer in microtitre plates. After appropriate incubation in 5 per cent CO2 atmosphere, the microtitre plate was examined for cytopathic effect. Cell detachment and shrinkage of Vero cells were recorded as toxic changes. RESULTS: All 36 environmental isolates demonstrated cytopathic effect of which 41.7, 50 and 8.3 per cent belonged to A. hydrophila, A. veronii biotype sobria and A. caviae respectively. INTERPRETATION & CONCLUSION: The results demonstrated the presence of potentially pathogenic environmental aeromonads in and around Vellore and they produced cytotoxin.Item Detection of opportunistic DNA viral infections by multiplex PCR among HIV infected individuals receiving care at a tertiary care hospital in South India.(2009-07) Sachithanandham, J; Ramamurthy, M; Kannangai, R; Daniel, H D; Abraham, O C; Rupali, P; Pulimood, S A; Abraham, A M; Sridharan, GPurpose: Opportunistic viral infections cause increased morbidity and mortality among human immunodeficiency virus (HIV) infected individuals, especially those who are not on antiretroviral treatment. Early diagnosis of these opportunistic viruses will be able to reduce the risk of disease progression with appropriate intervention. Materials and Methods: Multiplex PCR was attempted to detect the opportunistic herpes viruses (HSV-1, HSV-2, VZV, EBV, and CMV), adenovirus and polyoma viruses (JC and BK) in three cocktails of PCR reactions. Subsequently, all the viruses detected were quantitated by testing using monoplex real time PCR. Whole blood samples collected between 2006 and 2007 from 68 treatment naοve HIV-1 infected and 30 normal healthy individuals were tested for these eight viruses. Among the 68 HIV -1 infected individuals 35 had CD4+ T cell count less than or equal to 200 while the other 33 had greater than 200 CD4+ T cells. Results: Among the 68 HIV-1 infected individuals, 49 (72%) were positive for EBV, 5 (7%) samples were positive for CMV. All the five CMV positive individuals had CD4+ T cell count of less than or equal to 200 cells/µL. The mean EBV load among the individuals with a CD4+ T cells of less than or equal to 200 cells/µL was 3.88 log 10 while among those with greater than 200 CD4+ T cells it was 3.75 log 10 . The mean CMV load was 6.98 log 10. Three samples were positive for both CMV & EBV. None of the samples was positive for HSV-1, HSV-2, VZV, Adenovirus, JC and BK viruses. Conclusions: In our study, multiplex PCR based detection system was found useful in detecting opportunistic viruses in HIV infected individuals. Though EBV is the most prevalent opportunistic viral infection among HIV infected individuals, there was no significant association between EBV load, CD4+ T cell counts and HIV-1 virus load. CMV was seen in HIV infected individuals with low CD4+ T cell counts (less than 200 cells/μL).Item Detection of parvovirus B19 in selected high-risk patient groups & their phylogenetic & selection analysis(Indian Council of Medical Research, 2018-04) Vadivel, K; Mageshbabu, R; Sankar, S; Jain, A; Perumal, V; Srikanth, P; Ranjan, GA; Nair, A; Simoes, EA; Nandagopal, B; Sridharan, GBackground & objectives: Human parvovirus B19V (B19V) is known to be associated with erythema infectiosum commonly in children, aplastic crisis, especially in persons with underlying haemolytic disorders, hydrops fetalis in pregnancies and arthritis. This cross-sectional study was aimed to determine the presence of B19V infection in childhood febrile illnesses, association of B19V with arthropathies and in adult patients with end-stage renal disease (ESRD) on dialysis. The genetic diversity among the sequences was also analysed. Methods: A nested polymerase chain reaction (nPCR) assay was used for B19V DNA targeting VP1/VP2 region and used for testing 618 patients and 100 healthy controls. Phylogenetic analysis on nucleotide and amino acid sequences was carried out to compare our sequences with other Indian strains and global strains. Results: Among 618 samples tested, seven (1.13%) were found positive. The phylogenetic analysis revealed that all the seven sequences belonged to genotype 1 and showed low genetic diversity. The clustering pattern of seven sequences was similar both by nucleotide and by predicted amino acid sequences. The fixed effects likelihood analysis showed no positive or negatively selected sites. Interpretation & conclusions: Seven samples (4 from non-traumatic arthropathies, 2 from patients with ESRD and 1 from febrile illness patient) were found positive by nPCR. When our seven sequences were compared with global strains, the closest neighbour was other Indian strains followed by the Tunisian strains.Item Detection of virulence attributes of Burkholderia pseudomallei.(2004-03-30) Balaji, V; Jesudason, Mary V; Sridharan, G; Subramanian, KBACKGROUND & OBJECTIVES: Melioidosis caused by Burkholderia pseudomallei is an emerging disease in India. This study examined the toxin activity of bacteria-free culture filtrate in three different cell lines (cytotoxic assay) and its effect on Caenorhabditis elegans (nematode toxicity assay). Endotoxic activity of the viable bacteria was also studied in C. elegans (co-culture killing assay). METHODS: For toxin studies, serial doubling dilutions of unheated, heated crude and ultra filtrate of bacteria-free culture supernatants of B. pseudomallei were tested in 96-well microtitre plate containing confluent mono layers of McCoy, Hep-2 and HeLa cell lines. For the effects on C. elegans, the worms were exposed to heated and unheated bacteria-free culture supernatants in 24-well microtitre plate for 24h and then transferred to OP50 Escherichia coli lawn culture. The endotoxic activity of the live bacterium was studied by feeding the worms in the lawn culture of B. pseudomallei. RESULTS: All the clinical isolates (n=38) produced cytotoxic changes in all the cell lines. No difference was observed in the cytotoxicity of unheated, heated and ultra-filtered culture supernatant. The septicaemic isolates were observed to produce cytotoxic changes in high dilutions (1:160) of culture filtrate. None of the unheated and heated crude filtrate had deleterious effect on C. elegans, while all the live bacteria were found to be lethal to the nematode. INTERPRETATION & CONCLUSION: The culture supernatants, though produced cytopathic effect in various tissue cultures, failed to have any deleterious effect on the worms. However, live bacteria were lethal to the worms B. pseudomallei. Use of C. elegans model to detect virulence attributes of B. pseudomallei is recommended as an alternative to tissue culture methods as this can be carried out in laboratories where a tissue culture set up is not available.Item Diagnosis of typhoid fever by the detection of anti-LPS & anti-flagellin antibodies by ELISA.(1998-05-22) Jesudason, M V; Sridharan, G; Arulselvan, R; Babu, P G; John, T JIn a developing country like ours where typhoid fever is endemic and there are very few microbiology laboratories to provide diagnosis by culture, a search for non culture techniques for rapid and reliable diagnosis continues. The Widal test has a low sensitivity. We have attempted to adapt the well established enzyme linked immunosorbent assay (ELISA) technique to provide a serological test of better sensitivity and specificity. The ELISA described here to detect anti-LPS antibodies was found to have a sensitivity of 89.4 per cent and a specificity of 94.9 per cent. The sensitivity for the antiflagellin ELISA was 68.1 per cent and specificity was 97.4 per cent. The likelihood ratio was 17.5 for anti-LPS ELISA and 26.2 for the anti-flagellin ELISA. Considering the positivity of either one ELISA as diagnostic, the sensitivity was 93.6 per cent and specificity was 94.9 per cent.Item Double stranded DNA antibody estimation for the diagnosis of systematic lupus erythematosus [corrected](1993-07-01) Mohanalingam, U; Mathai, E; Sridharan, GThe usefulness of ds DNA antibody detection in the diagnosis of SLE in Indian population was evaluated. One hundred and fourteen sera were tested for antinuclear antibodies by indirect immunofluorescence and for ds DNA antibodies by ELISA. ELISA test has a specificity of 99% and a positive predictive value of 95% for the diagnosis of SLE.