Pcr detection of cytomegalovirus in umbilical cord blood

Nava Etminan taghva,1,* Mohammad hassan shahhosseiny,2 Fahimeh nemati mansour,3

1. Department Of Biotechnology, Faculty Of Advanced Sciences & technology, pharmaceutical Sciences Branch, Islamic Azad Uni
2. Department of Microbiology – shahr-e-Qods Branch – Islamic Azad University –Iranian Gene Fanavar Institute (IGF), Tehran
3. Department Of Biotechnology, Faculty Of Advanced Sciences & technology, pharmaceutical Sciences Branch, Islamic Azad Uni

Abstract

Introduction

Cytomegaloviruses (cmv) are herpes viruses are everywhere and common cause of human disease. the virus causes infections of the uterus and as a result, the transmission of the infection to the fetus, in the high incidence of congenital infections. for this reason, by designing this project, we are looking for cmv viral infection in the umbilical cord blood and since pcr and modern molecular techniques are a quick and accurate method, it help us to identify and diagnose viral infections early.

Methods

100 umbilical cord blood samples were prepared in sterile conditions from the operating room of mostafa khomeini hospital. dna was extracted from the umbilical cord blood ,samples using dng-plus method. the pcr test was optimized using specific primers for cmv based on the ß glycoprotein target gene. the pcr test was evaluated for limit of detection (lod) and specificity and then on extracted dna samples.

Results

The pcr test was optimized and the product of 257 bp was observed in 1.5% agarose gel electrophoresis. the lod was obtained in this study 100 copy/reaction, also primers no positive with other dna in the test of specificity. of 100 cord blood samples, 20% were contaminated by the presence of cmv virus.

Conclusion

Considering the role played by cmv in congenital infections and considering the high accuracy and high sensitivity of amplification molecular tests such as pcr and the results obtained in this study, it can be concluded that viral cmv is important for the occurrence of congenital infections.

Keywords

Cytomegalovirus, umbilical cord blood, detection, pcr