Veterinary Research Forum
Vet Res Forum

Design and implementation of a TaqMan® real-time PCR method for detection and quantification of bovine leukemia virus

Document Type : Original Article

Authors

Hassan Vahidi Emami 1
Arash Ghalyanchi Langeroudi 1
Seyed Masoud Hosseini 2
Hamideh Najafi 1

1 Department of Microbiology and Immunology, School of the Veterinary Medicine, University of Tehran, Tehran, Iran

2 Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran

https://doi.org/10.30466/vrf.2024.2016741.4084
Abstract
The bovine leukemia virus (BLV) is an important infectious agent transmitted from cattle to humans. It is considered one of the oncogenic viruses in breast cancer, so an accurate detection of this virus is important. The study aimed to design a specific and sensitive method based on TaqMan® real-time polymerase chain reaction (RT-PCR) for BLV detection. Probes and primers were designed using bioinformatics software for a 108 pairs region of the BLV tax gene. Criteria employed for determining analytical sensitivity were prepared using in-vitro RNA transcriptions. The National Center for Biotechnology Information (NCBI), basic local alignment search tool (BLAST) databases various viral panels and genomic samples from healthy individuals (Qom Province, Iran in 2023) were used to verify analytical specificity and clinical specificity, respectively. This method can measure a minimum of 10 copies of DNA and RNA mL-1. Moreover, the assay is linear in the range of 100 - 109 copies mL-1. By testing negative specimens, the method specificity was 100%. The reproducibility results of the reaction were examined at the intra- and inter-assay comparison. In fact, 10 technical replicates of each concentration of the control sample were analyzed in each working reaction. Due to the locally made kit, exact sensitivity and specificity, rapid analysis, and relatively low cost, as compared to commercial kits of other countries, the method introduced in the present study could be suitable for accurate detection of the BLV. Also, the TaqMan® real-time PCR method could be detected in cattle and human and before malignant changes of breast cancer which could reduce infection and breast cancer.

Keywords

Bovine leukemia virus
Breast cancer
Cattle
Real-time PCR
Subjects
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Veterinary Research Forum
Volume 15, Issue 8
August 2024
Pages 417-423

  • Receive Date 18 December 2023
  • Revise Date 19 February 2024
  • Accept Date 07 April 2024

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