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Development of Novel Multiplex PCR for Diagnosis of Co-infected Hemo-parasites in Cattle

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Title Development of Novel Multiplex PCR for Diagnosis of Co-infected Hemo-parasites in Cattle
Not Available
 
Creator Pankaj Kumar
Abhay Kumar
Kamal Sarma
Paresh Sharma
Rashmi R Kumari
Manish Kumar
 
Subject Anaplasma
Babesia
Co-infection
multiplex PCR
Theileria
 
Description Diagnostic Method
Background: A novel, rapid and specific multiplex polymerase chain reaction was developed to diagnose hemo-parasitic infection in
bovine blood co-infected with three of the most common hemo-parasites.
Methods: The diagnostic process relied on the detection of the three different bovine hemoparasites isolated from red blood cells
(RBCs) of cattle (N=30) by conventional Giemsa stained blood smear (GSBS) and confirmed by multiplex PCR. The multiplex PCR
system was used to diagnose GSBS positive blood samples (N=12) found infected or co-infected with hemoparasites. The designed
multiplex primer sets was attempted to amplify 205, 313 and 422 bp fragments of apocytochrome b, sporozoite and macroschizont 2
(spm2) and 16S rRNA gene for Babesia bigemina, Theileria annulata and Anaplasma marginale, respectively.
Result: This multiplex PCR was sensitive with the ability to detect the presence of 150 ng of genomic DNA. The primers used in this
multiplex PCR also showed highly specific amplification of specific gene fragments of each respective parasite. Comparing the two
detection methods revealed that 58.33% of specimens showed concordant diagnoses with both techniques. The specificity, positive
predictive value and kappa coefficient of the agreement was highest for diagnosis of B. bigemina and lowest for A. marginale. The
overall Kappa coefficient for diagnosis based on GSBS for multiple pathogens compared to multiplex PCR was 0.56, slightly behind
the threshold of 0.6 of agreement. Therefore, confirmation should always be based on PCR to rule out false positives due to differences
in subjective observations, stain particles and false negatives due to low parasitemia. The simplicity and rapidity of this specific
multiplex PCR method make it suitable for large-scale epidemiological studies and follow-up of drug treatments.
Not Available
 
Date 2021-09-23T07:12:19Z
2021-09-23T07:12:19Z
2021-09-09
 
Type Article
 
Identifier : Kumar, P., Kumar, A., Sarma, K., Sharma, P., Kumari, R.R., Kumar, M. (2021). Development of Novel Multiplex PCR for Diagnosis of Co-infected Hemo-parasites in Cattle. Indian Journal of Animal Research. DOI: 10.18805/IJAR.B-4695.
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/64601
 
Language English
 
Relation Not Available;
 
Publisher ARCC Karnal