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Journal code(s)
- E-ISSN Number: 2277-7695
- P-ISSN Number: 2349-8242
- CODEN Code: PIHNBQ
- ZDB-Number: 2663038-2
- IC Journal ID: 7725
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Vol. 11, Special Issue 7 (2022)
Environmental friendly rapid extraction method of nucleic acid for diagnosis of tropical theileriosis
Author(s):
Sumnil Marwaha, Basanti Brar, Vinod Kumar Jain and Minakshi Prasad
Sumnil Marwaha, Basanti Brar, Vinod Kumar Jain and Minakshi Prasad
Abstract:
Background: Tropical theileriosis is a haemoprotozoan disease causing huge economic losses in livestock sector. PCR is the most sensitive and specific for theileriosis. For PCR, DNA extraction is an important prerequisite step. A good DNA extraction method must be rapid, safe and yield good quality DNA. All of these criteria are met by DNA extraction method using Chelex-100. Chelex-100 is ionic resin that chelates ions like magnesium and other PCR inhibitors from sample yielding good quality and stable DNA that can be used for PCR.
Background: Tropical theileriosis is a haemoprotozoan disease causing huge economic losses in livestock sector. PCR is the most sensitive and specific for theileriosis. For PCR, DNA extraction is an important prerequisite step. A good DNA extraction method must be rapid, safe and yield good quality DNA. All of these criteria are met by DNA extraction method using Chelex-100. Chelex-100 is ionic resin that chelates ions like magnesium and other PCR inhibitors from sample yielding good quality and stable DNA that can be used for PCR.
Method: In present study, a method using Chelex-100 was standardized for DNA extraction from blood. A novel PCR assay was standardized for Theileria annulata using primers designed for TAMS1 gene. For comparison, DNA was extracted from 20 blood samples from theileriosis affected cows using both the Chelex-100 method and commercial kit method. Each DNA sample was used for detection of tropical theileriosis using standardized PCR assay.
Result: The average yield of DNA was 61.36 ng/µL and 99.7 ng/µL by Chelex-100 and commercial kit method, respectively. The amount of blood required for extraction was 25 µL and 200 µL in Chelex method and kit method, respectively. The DNA samples extracted by both method yielded positive results in PCR.
Pages: 1409-1412 | 465 Views 293 Downloads
How to cite this article:
Sumnil Marwaha, Basanti Brar, Vinod Kumar Jain and Minakshi Prasad. Environmental friendly rapid extraction method of nucleic acid for diagnosis of tropical theileriosis. The Pharma Innovation Journal. 2022; 11(7S): 1409-1412.
Journal code(s)
- E-ISSN Number: 2277-7695
- P-ISSN Number: 2349-8242
- CODEN Code: PIHNBQ
- ZDB-Number: 2663038-2
- IC Journal ID: 7725
Main Menu
Special Issue
Copyright Form
Identifier
The Pharma Innovation Journal
