Repository of Research and Investigative Information

Repository of Research and Investigative Information

دانشگاه علوم پزشکی و خدمات بهداشتی درمانی زنجان

Simultaneous Detection and Genotype Determination of HSV 1 and 2 by Real-time PCR Using Melting Curve Analysis and a Unique Pair of Primers

(2017) Simultaneous Detection and Genotype Determination of HSV 1 and 2 by Real-time PCR Using Melting Curve Analysis and a Unique Pair of Primers. Applied immunohistochemistry & molecular morphology : AIMM. pp. 139-143. ISSN 1533-4058 (Electronic) 1533-4058 (Linking)

Full text not available from this repository. (Request a copy)

Official URL: http://www.ncbi.nlm.nih.gov/pubmed/26709736

Abstract

Herpes simplex virus (HSV) is a human pathogen that causes different pathologic manifestations. Rapid and feasible detection and discrimination methods for HSV genotyping is a challenge in clinical laboratories, especially in children suffering from herpetic encephalitis. A quantitative real-time polymerase chain reaction (PCR)-based genotyping assay using SYBR Green I was established. We designed only 1 pair of primer for HSV 1 and 2, targeting thymidine kinase gene conserved region. HSV genotypes were determined by PCR using melting curve analysis with LightCycler. Different HSV genotypes were successfully detected in all clinical samples. The melting temperature for HSV 1 and 2 was 85.5+/-0.78 degrees C and 89+/-0.53 degrees C, respectively. These 2 genotypes were completely distinguished by means of the accurate melting assay. Importantly, detection was reliably performed within only 1 hour. The assay had no cross-reactivity across species, an excellent dynamic range from 10 to 10 copies per reaction, a good intra-assay and interassay reproducibility, and a detection limit of a single copy per reaction. Our homebrew designed and validated quantitative real-time PCR followed by a melting curve analysis provided a rapid and convenient screening test for differential identification of HSV genotypes 1 and 2. We recommend the large-scale application of this method for HSV 1 and 2 detection.

Item Type: Article
Keywords: DNA Primers Genes, Viral Herpes Simplex/virology Herpesvirus 1, Human/enzymology/*genetics Herpesvirus 2, Human/enzymology/*genetics Humans Limit of Detection Real-Time Polymerase Chain Reaction/*methods Reproducibility of Results Thymidine Kinase/genetics
Page Range: pp. 139-143
Journal or Publication Title: Applied immunohistochemistry & molecular morphology : AIMM
Abstract and Indexing: ISI, Pubmed, Scopus
Quartile : Q2
Volume: 25
Number: 2
Identification Number: https://doi.org/10.1097/PAI.0000000000000282 http://apps.webofknowledge.com/full_record.do?product=WOS&search_mode=GeneralSearch&qid=66&SID=C48LsSa76YoWspN8AAK&page=1&doc=4
ISSN: 1533-4058 (Electronic) 1533-4058 (Linking)
Depositing User: خانم مریم زرقانی
URI: http://repository.zums.ac.ir/id/eprint/346

Actions (login required)

View Item View Item