Abstract
Aim: To clone the virulent gene LipL32 of Leptospira interrogans serovar Autumnalis and to analyze the sequence with LipL32 gene of other pathogenic serovars of Leptopsira. Materials and Methods: Leptospira interrogans serovar Autumnalis procured from Leptospira research laboratory, Chennai was used in the study. Polymerase chain reaction (PCR) was carried out for amplifying LipL32 gene using the reported primers of Leptospira Kirschnerii. The PCR product was cloned into TA cloning vector and the vector was transformed into E.Coli DH5a cells. The plasmid was isolated from E.Coli and sent for sequencing with universal primers. The sequence was submitted in genbank with accession number JQ861883. Results: The PCR product revealed an amplicon of 790 bp. The LipL32 gene sequence of Leptospira interrogans serovar Autumnalis showed 99 % similarity with most of the pathogenic Leptospires. Conclusions: LipL32 gene of Leptospira is highly conserved in most of the pathogenic Leptospires. The study concludes that this gene could be used as a target for the diagnosis of leptospirosis in animals and humans and could be tested as an important candidate antigen for vaccine production.
Highlights
Leptospirosis has re-emerged as an important zoonotic disease in India and is endemic in most of the southern states like Kerala, Tamil Nadu and certain parts of Andhra Pradesh [1,2,3]
The study concludes that this gene could be used as a target for the diagnosis of leptospirosis in animals and humans and could be tested as an important candidate antigen for vaccine production
LipL32 was consistently observed in all pathogenic leptospires and was observed to be the immunodominant protein antigen recognized by the human humoral response during natural infections
Summary
Leptospirosis has re-emerged as an important zoonotic disease in India and is endemic in most of the southern states like Kerala, Tamil Nadu and certain parts of Andhra Pradesh [1,2,3]. The tests employing outer membrane proteins (OMPs) showed high sensitivity and specificity [4,5]. LipL32 was consistently observed in all pathogenic leptospires and was observed to be the immunodominant protein antigen recognized by the human humoral response during natural infections. This protein could be useful as a marker of infection for laboratory case confirmation in the field and in differentiating leptospirosis from other causes of acute febrile illness [7]. Leptospiral protein p32 is the most immunodominant protein and could be employed for the development of novel diagnostic assays [8]. The present study was carried out to clone and sequence the LipL32 gene of Leptospira interrogans serovar Autumnalis and to analyze the sequence relationship with other pathogenic leptospires
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