Madhavaram Milk Colony, India
Madhavaram Milk Colony, India

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Ram S.,L and crobiology Research Center | Vimalin J.M.,L and crobiology Research Center | Jambulingam M.,L and crobiology Research Center | Tiru V.,Sundaram Medical Foundation | And 2 more authors.
Malaysian Journal of Microbiology | Year: 2012

Aim: Isolation, dark field detection and microscopic agglutination test (MAT) are considered"gold standard" tests for diagnosis of Leptospirosis. Several PCR assays are reported but very few have been evaluated for detection of Leptospirosis. Therefore, this study was undertaken. This study aims to design and standardize polymerase chain reaction (PCR) -based DNA sequencing technique for the detection of pathogenic Leptospira from peripheral blood of patients clinically diagnosed with septicemia. Methodology and Results: Two hundred and seven (207) blood samples from patients were diagnosed with septicemia which includes 100 bacterial (other than Leptospira) culture positive and 107 bacterial culture negative samples were studied. Primers for Nested PCR targeting LipL32 gene of Leptospira interrogans were designed and the specificity of primers was tested against serum samples positive/negative by either MAT or dark field microscopy. PCR amplified products were further confirmed by DNA sequencing. The standardized nPCR was sensitive and specific to Leptospira interrogans. Twenty-one (21%) out of 100 culture positive blood samples, three (2.8%) out of 107 culture negative samples showed nPCR positivity and were confirmed as Leptospira interrogans by DNA sequencing (p<0.001). A sensitive nPCR specific to Leptospira interrogans was developed. Conclusion, significance and impact of study: The p value (<0.001) signifies that Leptospira is commonly associated with other bacteria circulating in blood indicating that a decreased immune status is created primarily by a bacterium with enhanced possibility of development of Leptospiral infection probably be of an endogenous origin.

Selvaraj J.,Central University of Costa Rica | Murali Manohar B.,Central University of Costa Rica | Govindarajan R.,Leptospirosis Research Laboratory | Jayakumar V.,Leptospirosis Research Laboratory | And 2 more authors.
Buffalo Bulletin | Year: 2010

Seroprevalence of leptospirosis in shebuffaloes in Chennai, India were detected by microscopic agglutination test. Leptospirosis was detected in 88 percent of 125 sera samples tested. The most prevalent serogroup observed was Pomona (54.4 percent). Highest titre of 6400 was detected with the serogroup Pomona (0.4 percent).

Ramesh S.,Madras Veterinary College | Jayakumar V.,Leptospirosis Research Laboratory | Murali Manohar B.,Leptospirosis Research Laboratory
Asian Journal of Microbiology, Biotechnology and Environmental Sciences | Year: 2011

Plants have been a valuable source of natural products for maintaining human and animal health, especially in the last decade with more intensive studies for natural therapies. Leptospirosis is a zoonotic disease, occurs mostly in tropical and subtropical developing countries and is caused by spirochetes of genus Leptospira. Treatment of leptospirosis includes antibiotics and supportive therapy to cure and to improve the health of the patients. Hence, holistic approaches are needed to treat the patients. Many herbal extracts exert not only antibiotic activity but also exhibit other activities like immune enhancer etc. In this study, we had used 11 herbal extracts to evaluate their anti-leptospiral activities and only Andrographis paniculata extracts had shown this property. The MIC 90 and MIC 50 for Andrographis paniculata extract were 62.5μg and 31μg respectively for all leptospiral serovar tested. From our study, it has been inferred that the Andrographis paniculata extract hold good promise as it is proved to have anti-leptospiral activity against all species of Leptospira serovars tested. The wide tissue and organ distribution and the hepato-protective, immune stimulating action of Andrographis paniculata with anti-leptospiral activity makes it an ideal candidate for the prevention and treatment of leptospirosis. © Global Science Publications.

Anandachitra M.,Leptospirosis Research Laboratory | Jayakumar V.,Leptospirosis Research Laboratory | Murali Manohar B.,Leptospirosis Research Laboratory
Asian Journal of Microbiology, Biotechnology and Environmental Sciences | Year: 2011

Novel leptospiral antigens like Lig A, B and Lsa21 were identified through screening of genomic DNA expression libraries with patient and infected animal sera and appeared to be preferentially expressed during host infection. Hence, the present study was attempted to detect these immunogenic proteins in field isolated leptospires and compare their sequences with the published reference strains. Field isolates of leptospires were identified as L.interrogans serovar australis and L.interrogans serovar hebdomadis by RAPD-PCR and using reference serum. Lig B and Lsa 21 genes were amplified in both field isolated leptospires with the amplicon size of 1100 bp and 540 bp respectively. LigB nucleotide sequences were having 93-100% identity with LigB sequences of several L.interrogans serovars and strains and only 91-92% identity with L.krischneri serovars and strains. Lsa21 nucleotide sequences are having 100% identity with query coverage of 97-100% with the predicted hypothetical protein sequences of L.interrogans serovar copenhageni and Lai for which complete genome sequences are available. From this study, we interpret that Lsa21 is more conserved than LigB protein in leptospires. Predicted amino acid sequences of both sequences revealed that they consist of mainly beta-pleated sheet with no or less alpha helical secondary structure. These predicted proteins are more hydrophilic with less hydrophobic regions and are more antigenic. This study indirectly will help in the usefulness of these recombinant protein based serodiagnosis and vaccine research. © Global Science Publications.

Magudeswaran S.K.,Madras Veterinary College | Parthiban M.,Madras Veterinary College | Saranya R.,Madras Veterinary College | Senthilkumar S.,Madras Veterinary College | Ravikumar G.,Leptospirosis Research Laboratory
Indian Journal of Biotechnology | Year: 2014

The present study was undertaken to compare the cross reactive potentiality of expressed recombinant protein (LipL41) with lipopolysaccharide (LPS) antigen of leptospira against different serovars of leptospira. Antigens from both recombinant LipL41 and the isolated LPS from Leptospira icterohaemorrhagiae were coated in latex beads and used to test the human sera samples of different serovars of leptospira using field based latex agglutination test. Recombinant antigen showed strong cross reaction with different serovars of leptospira, whereas LPS antigen showed very mild cross reaction with few serovars. This clearly indicates that outer membrane protein cross reacted with heterologous serovars and can be used as effective diagnostic agent as well as candidate antigen for development of vaccine for leptospira.

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