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Evaluation of Consensus-Degenerate Hybrid Oligonucleotide Priner (CODEHOP) and herpesvirus panel to detection of human herpesviruses in plasma samples of patients transplanted hematopoietic stem cells (HSCT)

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Author(s):
Talita Cristina Dellariva
Total Authors: 1
Document type: Master's Dissertation
Defense date:
Abstract

Introduction: Human herpesviruses are an important group can cause infections in immunocompromised patients and transplanted in general. Specifically in patients undergoing transplantation of hematopoietic stem cells, the members of the Herpesviridae family have been associated with the occurrence of high rates of morbidity and mortality. Currently, molecular methods of detection and quantification of infectious agents have been widely implemented in routine laboratories in order to minimize the impacts and prevent clinical manifestations caused by herpesviruses. Objectives: This study was standardized and optimized techniques Consensus-Degenerate Hybrid Oligonucleotide Primer in Nested-PCR format (CODEHOP NPCR) for the detection of human herpesvirus subfamilies and Human Herpesvirus Panel in nested PCR Touchdown format (TD NPCR) to detect the genome of the species of herpesviruses in plasma samples of patients transplanted hematopoietic stem cells and, subsequently, both techniques were evaluated from the comparison with the NPCR technique already established in the laboratory, used as a reference pattern. Results: From the optimization of CODEHOP NPCR technique was able to detect the three subfamilies of herpesviruses in positive control samples and the samples studied, and the comparison with the NPCR used as reference standard indicated that there is a moderate correlation between the technical and accuracy above 80%. Through the TD NPCR, all species could be amplified by this technique optimized and, the comparison with the reference standard technique, considerable excellent concordance was obtained for the different species, whereas the accuracy remained above 90%. By comparing the positivity median days after transplantation there was no significant difference between the techniques, indicating that can be applied for early detection of active infection by Human Herpesvírus. Conclusions: The techniques have increased the possibilities for molecular studies to be conducted in clinical samples in the laboratory and the early identification of patients at higher risk of developing disease herpesvirus after transplantation, favoring the use of antiviral drugs only in these cases (AU)

FAPESP's process: 14/16244-9 - Evaluation of Consensus-Degenerate Hybrid Oligonucleotide Primer (CODEHOP) and herpesvirus viral panel (panherpes) in detection of human herpesvirus in patients Transplanted Hematopoietic Stem Cells (HSCT)
Grantee:Talita Cristina Dellariva
Support type: Scholarships in Brazil - Master