Open Access
Research
(Published
online: 30-09-2016)
21.
Comparative evaluation of polymerase chain
reaction assay with microscopy for detection of asymptomatic
carrier state of theileriosis in a herd of crossbred cattle -
Gaurav Charaya, N. K. Rakha, Sushila Maan, Aman Kumar, Tarun
Kumar and Ricky Jhambh
Veterinary World, 9(9): 1039-1042
doi:
10.14202/vetworld.2016.1039-1042
Gaurav Charaya:
Department of Veterinary Medicine, Lala Lajpat Rai University of
Veterinary and Animal Sciences, Hisar, Haryana, India;
gcharaya9@gmail.com
N. K. Rakha:
Department of Veterinary Medicine, Lala Lajpat Rai University of
Veterinary and Animal Sciences, Hisar, Haryana, India; drrakha@gmail.com
Sushila Maan:
Department of Animal Biotechnology, Lala Lajpat Rai University
of Veterinary and Animal Sciences, Hisar, Haryana, India;
sushilamaan105@gmail.com
Aman Kumar:
Department of Animal Biotechnology, Lala Lajpat Rai University
of Veterinary and Animal Sciences, Hisar, Haryana, India;
aman.abt@luvas.edu.in
Tarun Kumar:
Teaching Veterinary Clinical Complex, Lala Lajpat Rai University
of Veterinary and Animal Sciences, Hisar, Haryana, India;
tarunvet@gmail.com
Ricky Jhambh:
Department of Veterinary Medicine, Lala Lajpat Rai University of
Veterinary and Animal Sciences, Hisar, Haryana, India;
jhambricky@gmail.com
Received: 05-05-2016, Accepted: 18-08-2016, Published online:
30-09-2016
Corresponding author:
N. K. Rakha, e-mail: drrakha@gmail.com
Citation:
Charaya G, Rakha NK, Maan S, Kumar A, Kumar T, Jhambh R (2016)
Comparative evaluation of polymerase chain reaction assay with
microscopy for detection of asymptomatic carrier state of
theileriosis in a herd of crossbred cattle,
Veterinary World, 9(9):
1039-1042.
Abstract
Aim:
This study aims to develop and to standardize a polymerase chain
reaction (PCR) assay that will diagnose clinical as well as
carrier state of the disease and to compare the results with
conventional microscopy technique.
Materials and Methods:
A herd of crossbred cattle with the previous history of
theileriosis in village Lahli, district Rohtak, Haryana, was
selected for this study. A total of 29 blood samples were
collected randomly from cows including five clinically ill
cattle. Blood smears from all animals and lymph node biopsy
smears from animal with swollen lymph nodes were examined
microscopically after conventional Giemsa staining. Phenol
chloroform isoamyl alcohol method was used for extracting DNA
from blood. Previously published primers targeting
cytochrome b
gene sequence of
Theileria annulata
were used in the PCR assay that was standardized to use in the
laboratory.
Results:
Out of 29 samples tested,18 (62.06%) were found positive for
theileriosis by PCR assay, whereas only 10 (34.48%) samples were
detected positive by conventional microscopic technique using
Giemsa staining method.
Conclusions:
On the basis results of comparative studies, it can be concluded
that PCR assay is a more sensitive than microscopic examination
for detection of theileriosis. This can be attributed to the
ability of PCR assay to detect small amounts of genomic DNA of
T. annulata
or low parasitemia in cows. Therefore, PCR assay can serve as a
more sensitive tool to detect
Theileria
for detection of theileriosis even in asymptomatic carrier
cattle which is important for the implementation of successful
control programs.
Keywords:
Carrier state,
cytochrome b,
Giemsa staining, polymerase chain reaction assay, theileriosis,
Theileria annulata.
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