International Journal of Pharma and Bio Sciences
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editorijpbs@rediffmail.com (or) editorofijpbs@yahoo.com (or) prasmol@rediffmail.com
10.22376/ijpbs.2019.10.1.p1-12
Volume 3 Issue 3
2012 (July - September)
Pcr-Sscp: A Tool For Molecular Diagnosis Of Leptospirosis
Leptospirosis, a disease with protean manifestations, is caused by lessThan i greaterThan Leptospira lessThan /i greaterThan species. Early and reliable identification of lessThan i greaterThan Leptospira lessThan /i greaterThan serovars remains to be a challenge due to diversity at serovar level. The potential of polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) was assessed for detection of sequence variation in pathogenic specific 631 bp fragment of 16S rDNA of lessThan i greaterThan Leptospira lessThan /i greaterThan for serovar typing. Standardization of this technique on reference serovars of lessThan i greaterThan L. interrogans lessThan /i greaterThan (Canicola, Pomona and Icterohaemorrhagiae) and of lessThan i greaterThan L. borgpetersenii lessThan /i greaterThan (Ballum, Tarassovi and Javanica) yielded distinct SSCP profiles. Employing this technique 15 isolates could be characterized to serovars Canicola, Icterohaemorrhagiae, Pomona and Javanica. Similar results were obtained when a panel of monoclonal antibodies in ELISA typed these isolates. In addition, this technique was applied on blood samples of human leptospirosis for direct identification and characterization of lessThan i greaterThan Leptospira lessThan /i greaterThan serovars. Five of 50 samples were characterized as serovar Tarassovi. DNA sequencing of 631bp amplicons of these samples validated the results obtained with PCR-SSCP. This PCR-SSCP was therefore, able to identify and characterize the lessThan i greaterThan Leptospira lessThan /i greaterThan to serovar level obviating the need for isolation of leptospires before testing.
Madhurima De Roy,K.Thavachelvam,H.V. Batra2 And U. Tuteja
Leptospira spp., PCR, SSCP
179-186