Molecular detection of Anaplasma marginale infection in carrier cattle.
Anaplasma marginale infections are responsible for widespread morbidity and mortality particularly in crossbred and exotic breeds of cattle in the tropics and subtropics. In the present study, a semi-nested PCR assay was utilized for the detection of A. marginale infection in carrier cattle from different areas of Punjab state, India. An amplicon of 458 bp specific for msp5 of A. marginale was detected in 45.2% of blood samples when subjected to primary PCR assay against the routine blood smear examination, which revealed inclusion bodies in only 12.5% of samples. Semi-nested PCR employing product of samples negative by primary PCR produced the amplicons of desired size (345 bp) in 51% (29/57) of samples indicating that nested PCR, when coupled with primary PCR, resulted in increased sensitivity in detection of A. marginale infection in carrier cattle from 45.2 to 73.1%. These results suggest higher threshold detection limits of the nested PCR over the conventional technique used for diagnosis of anaplasmosis which is ideally suited for diagnosis of carrier cases which act as nidus for the spread of the infection to the susceptible stock in endemic areas.
Department of Veterinary Parasitology, College of Veterinary Science, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab 141004, India. drharkiratsingh@hotmail.com
Harkirat
H
Singh
Jyoti
Manjurul
M
Haque
Nirbhay K
NK
Singh
Shitanshu S
SS
Rath
D042322:Anaplasma marginale / Q000235:genetics / Q000302:isolation & purification*; D000712:Anaplasmosis / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D002353:Carrier State / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology; D004269:DNA, Bacterial / Q000235:genetics; D007194:India / Q000453:epidemiology; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary
D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; C111578:major surface protein 5, Anaplasma