Question

Comparison of primer sets for detection of theileria annulata

Answer 1

Explanation:

140 native cows with the mean age of more than one year were selected randomly and their age and sex were registered in the related papers. Primarily, a thin layer smear was prepared from their ear sublime vein blood and was fixed with methanol and then it stained with Gimsa dye. Also, 9 mL blood from the jagular vein of the same cow was taken and collected in autoclaved tubes (containing 1 mL of 3.2% buffered citrate solution) conversely for extracting their DNA for PCR. Two primers were used: N516/N517 belonged to a huge gene (30 kDa) which is responsible for coding the surface antigen of Theileria annulata merozoite. By detection of 140 prepared blood smear with light microscope and lens 100, piroplasmic forms of Theileria annulata were seen in 12 smear (8.57%). By PCR method, 56 DNA of Theileria annulata was separated from 140 samples (40%). Statistical comparison of PCR and smear methods in diagnosing of Theileria annulata carriers can explain a significant difference between these two methods (p<0.05). The results of this study shows that sensitivity and accuracy of PCR method in diagnosing of Theileria annulata carriers is more than common method of smear preparation and can be used in epidemiological studies for the sake of controlling, prevention and determining of immunological condition of cows in region.

I hope this will help u

please mark me as brainliest

Answer 2

Explanation:

The polymorphic nature of Theileria annulata merozoite surface antigen (TAMS 1) attributes to limitation in PCR based detection of various T. annulata genotypes present in different geographical domains across the globe. Multiple reports of failure of detection of tropical theileriosis using classical N516/517 primer set in the studied area were noticed. Hence, three single PCR protocols using N516/517, TAMS F/R and NTA F/R primer sets encoding different portions of TAMS 1 gene and two nested protocols, using combinations of these three primers, were compared to find out the most suitable primer set for diagnosis of calf theileriosis in studied area. The studied area constitutes the semi-arid theileriosis endemic area of Northern India. The various PCR protocols were tested on 75 clinically confirmed cases of calf theileriosis. Alongside, 25 confirmed theileriosis negative blood samples and DNA of other haemoprotozoa were also tested for specificity of these primer sets. Results revealed that the primer set NTA F/R to be more suitable in detecting the circulating T. annulata genotypes in the studied area in comparison to the classical N516/517 primer set. None of the primers gave false positive amplification with negative samples and/or DNA of other haemoprotozoa.