Standardization and validation of simple PCR, duplex PCR and RAPD in comparison to blood smear examination for diagnosing bovine tropical theileriosis.
Bovine Tropical Theileriosis (BTT) is an important vector-borne protozoan disease that imposing serious constraints on the health and productivity of domestic cattle. It is matter of common fact that following recovery from primary infection, cattle become persistent carriers and act as reservoirs of infection thereby, playing a critical role in disease epidemiology. The present study describes the comparative diagnostic efficiency of simplex PCR, duplex PCR and RAPD assays for detection of Theileria annulata in cattle. An optimized simple PCR and duplex PCR assay were established using TAMS F/R as primer sets encoding for 721 bp amplicon alongside a RAPD with arbitrary primer coding for 963 bp product of T. annulata. The simple PCR and duplex PCR detected pathogen with almost same level of sensitivity, irrespective of whether its DNA was amplified in isolation or together with DNA of another pathogen without nonspecific amplifications. RAPD failed to give comparable results and suffered from limitations of sensitivity as well as specificity. The developed assays may be seen as a good tool for epidemiological studies aiming at assessing the burden of chronic infections and improving control of the associated diseases in endemic regions.
College of Veterinary Sciences and Animal Husbandry, UP Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura 281001, India. Electronic address: viks.sudan@gmail.com.
Vikrant
V
Sudan
College of Veterinary Sciences and Animal Husbandry, UP Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura 281001, India.
Daya
D
Shanker
College of Veterinary Sciences and Animal Husbandry, UP Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura 281001, India.
Amit
A
Jaiswal
College of Veterinary Sciences and Animal Husbandry, UP Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura 281001, India.
Amit
A
Singh
College of Veterinary Sciences and Animal Husbandry, UP Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura 281001, India.
Vijay
V
Pandey
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000097:blood / Q000175:diagnosis* / Q000469:parasitology; D016054:DNA, Protozoan / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016794:Theileria / Q000235:genetics* / Q000502:physiology; D013801:Theileriasis / Q000097:blood / Q000175:diagnosis* / Q000469:parasitology
D016054:DNA, Protozoan
Cattle; Duplex PCR; RAPD; Simple PCR; Theileria annulata