EVALUATION OF MOLECULAR BEACON FLUO RESCENT PROBE FOR LABORATORY DIAGNOSIS OF CONGENITAL CYTOMEGALOVIRUS (CMV) INFECTION

G. Orrù

First

Methodology

;P. Melis;F. Puggioni;M. Puddu;G. Ottonello;V. Fanos

Second

Member of the Collaboration Group

;F. Coghe

2017-01-01

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Abstract

INTRODUCTION Cytomegalovirus is a species of DNA virus that belongs to Herpesviridae family and represents the leading cause of congenital infection worldwide. It occurs in 0.2% to 2% of live births and 1/5 of these could present long-term health problems. Infected patients spread the virus in cervical mucus, feces, breast milk, saliva even if viral detection of urine has been considered the most reliable method for diagnosis of CMV in newborn. Rapid and accurate laboratory diagnosis of CMV is a crucial point to support the right therapy and proper management of the infection. In this context, the molecular approach by real-time PCR method is a widely available procedure for viral detention/ quantification. This procedure is traditionally based on linear molecular probes (SYBR Green, TaqMan) able to issue fluorescence during the DNA amplification, although different authors reported technical problems in specificity and reproducibility by using these fluorescent probes. In other cases, these molecules require an expensive laboratory equipment, not suitable for example in developing countries. The objective of the present study was to compare traditional PCR TaqMan kit for CMV with a circular probe called Molecular Beacon (MB). The MBs are hairpin-shaped oligonucleotide probes, and have the property of fluorescing upon hybridization with a specific DNA sequence (Fig. 1). Their hairpin shape increases target specificity providing a lower background signal compared to that found in other linear probes previously cited.