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Exp Parasitol 2004 Mar-Apr;106(3-4):110-8

Arbitrary-primed PCR for genomic fingerprinting and identification of differentially regulated genes in Indian isolates of Leishmania donovani.

Sreenivas G, Singh R, Selvapandiyan A, Negi NS, Nakhasi HL, Salotra P

Salotra P, Safdarjang Hosp, ICMR, Inst Pathol, New Delhi 110029, India Safdarjang Hosp, ICMR, Inst Pathol, New Delhi 110029, India US FDA, CBER, OBRR, Div Emerging & Transfus Transmitted Dis, Bethesda, MD 20892 USA Safdarjang Hosp, Dept Med, New Delhi 110029, India


The arbitrary-primed PCR (AP-PCR) technique was employed with the twin goals of identifying genetic polymorphisms within the Indian isolates and to identify differentially expressed gene sequences. The parasite isolates from Indian Kala-azar patients could be differentiated from Leishmania donovani isolates from distinct geographic regions. Moreover, differences within the Indian isolates could also be identified. A majority (17/19) of the Indian isolates gave identical AP-PCR pattern, while two isolates gave consistently divergent pattern. The distinctive AP-PCR fragments obtained with Indian isolates were used as probes in Northern blot analysis. Three such fragments were found to represent transcribed sequences that were differentially expressed in the two stages of the parasite. These sequences led to cloning and characterization of Leishmania Centrin gene and a novel gene termed A-1 that is over-expressed in amastigote stage of the parasite. The study demonstrates the utility of random genome sampling methods in genomic fingerprinting and in identifying differentially transcribed sequences that could potentially contribute to parasite virulence.

Category: Journal Article, Peer
PubMed ID: #15172218
Includes FDA Authors from Scientific Area(s): Biologics
Entry Created: 2011-10-04 Entry Last Modified: 2012-08-29