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Articles by P. Thomas
Total Records ( 3 ) for P. Thomas
  K. Karthik , R. Rathore , P. Thomas , A. Elamurugan , T.R. Arun and K. Dhama
  Brucella abortus, one of the major pathogen causing abortions in cattle worldwide and a zoonotic agent, need to be detected earlier in order to prevent its spread among animals. The present study aimed at to know the prevalence of B. abortus in cattle population of three states (Uttar Pradesh, Uttarakhand and Tamil Nadu) of India by serological (Rose Bengal Plate Test (RBPT) and Serum Tube Agglutination Test (STAT)) and molecular (polymerase chain reaction) detection in sera samples and whole blood (n = 370), respectively. Out of a total of 370 sera samples, 61 (16.49%) were positive by RBPT and 59 (15.94%) by STAT. Screening of the whole blood samples by genus specific bcsp31 gene based PCR as well as species specific IS711 gene based PCR revealed that 56 (15.13%) samples were positive for brucellosis. None of the serologically negative sample showed positivity by PCR; however few positive samples were tested negative by PCR. Sensitivity and specificity of PCR compared with RBPT was 100 and 92.4% while with STAT these were 100 and 95.16%, respectively. Results are promising that whole blood can be used for studying the molecular epidemiology of B. abortus in cattle and particularly detecting the active phase of infection and PCR can be well adopted as a valuable test for mass screening of animals for this purpose. The present study adds to the prevalence data available regarding to B. abortus infection in cattle population and highlights the usefulness and advantages of molecular tool of PCR over serological tests.
  P. Thomas , T.R. Arun , K. Karthik , P.V. Berin , M. Asok Kumar , Neetu Singh , J. Usharani , M. Palanivelu , S.K. Gupta , K. Dhama and K.N. Viswas
  Necrotic enteritis, caused by Clostridium perfringens, is an important bacterial disease of poultry. A suspected case of necrotic enteritis was presented for necropsy from an Indian Kadaknath Fowl flock showing diarrhea and progressive debility. Gross examination revealed necrotic to ulcerative lesions in intestine. The organism was isolated from the intestinal contents, tissue and liver under anerobic conditions. The cultural characteristics and Gram staining were suggestive of C. perfringens. The sequencing of 16s rRNA gene confirmed the isolate as C. perfringens and which was well differentiated from other clostridia associated with avian intestinal tract. This study demonstrates that 16s rRNA gene sequencing can provide rapid and confirmatory identification of C. perfringens. Further, Multiplex Polymerase Chain Reaction (mPCR) was performed for toxinotyping and isolate was found to be positive for α toxin (cpa) and β2 toxin (cpb2), a feature of C. perfringens type A isolates. As some recent studies have highlighted the involvement of NetB toxin in pathogenesis, therefore, PCR was carried out to find the presence of this toxin, the isolate was found to be negative for netB gene. This study emphasizes the molecular characterization and toxinotyping as a rapid tool for detection of C. perfringens from suspected necrotic enteritis cases. Very few reports regarding molecular characterization are available from India, hence it adds to the available data on this important poultry pathogen. Further investigations are required to understand the exact role of NetB toxin in pathogenesis as various studies including the current one reports NetB negative strains involved in necrotic enteritis.
  T.R. Arun , R. Rana , P. Singh , P. Choudhuri , V.P. Singh , P. Thomas , V. Rekha , K. Nehra , J. Usharani and K. Dhama
  A gold nanoparticle based lateral flow assay was developed for rapid serodiagnosis of contagious agalactia, an economically important mycoplasmal disease of small ruminants. Sonicated antigen of Mycoplasma agalactiae was used as the test reagent that was immobilized on nitrocellulose membrane along with the control line of goat IgG. The detection reagent, gold nanoparticle conjugated with anti-goat antibody was dried on the conjugate pad. During the assay, specific antibodies against M. agalactiae in the test serum that combined with the detection reagent were captured in the test line and detected visually by the development of a red line on nitrocellulose membrane. The gold conjugate captured in control line produced a red line regardless of the presence of specific antibodies that served as a procedural control. Serum samples collected from an experimentally infected goat were tested with the lateral flow assay and antibodies were detected from 9th day of infection and the assay was also evaluated using 100 goat sera samples. This is the first report regarding development of a gold nanoparticle based lateral flow assay for rapid diagnosis of contagious agalactia in goats. This study suggests that current lateral flow assay can be used as a user friendly diagnostic in laboratories lacking specialized equipments as well as for point of care diagnosis of contagious agalactia.
 
 
 
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