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Veterinary Research Forum، جلد ۱۵، شماره ۸، صفحات ۴۱۷-۴۲۳
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عنوان انگلیسی Design and implementation of a TaqMan® real-time PCR method for detection and quantification of bovine leukemia virus
چکیده انگلیسی مقاله 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.
کلیدواژه‌های انگلیسی مقاله Bovine leukemia virus,Breast cancer,Cattle,Real-time PCR
نویسندگان مقاله Hassan Vahidi Emami |
Department of Microbiology and Immunology, School of the Veterinary Medicine, University of Tehran, Tehran, Iran

Arash Ghalyanchi Langeroudi |
Department of Microbiology and Immunology, School of the Veterinary Medicine, University of Tehran, Tehran, Iran

Seyed Masoud Hosseini |
Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran

Hamideh Najafi |
Department of Microbiology and Immunology, School of the Veterinary Medicine, University of Tehran, Tehran, Iran

نشانی اینترنتی https://vrf.iranjournals.ir/article_714438_cd6e2986cd7f2a18171a3ba191039112.pdf
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