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| Author(s): |
Renato Souza Oliveira
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Faculdade de Ciências Médicas |
| Defense date: | 2015-08-20 |
| Examining board members: |
Sandra Helena Alves Bonon;
Celso Francisco Hernandes Granato;
Magnun Nueldo Nunes dos Santos
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| Advisor: | Sandra Helena Alves Bonon |
| Abstract | |
Introduction: Human herpesviruses (HHV) are the major causes of infections in transplant and Immunocompromised patients in general. High levels of HHV-6 are associated with increased mortality, graft versus host disease (GVHD), HCMV disease and encephalitis in hematopoietic stem cell transplant (HSCT) patients. HHV-7 can cause fever, rash and is possible cofactor for HCMV disease. Currently, molecular methods for detection and quantification of infectious agents are being used in routine in order to provide early treatment with appropriate antiviral (preemptive therapy), mainly for HCMV. The Real Time Polymerase Chain Reaction (qPCR) is one of the modern options to be used in the monitoring of transplant patients. One advantage of this technique is the possibility of using plasma from neutropenic patients, where this situation often affect the realization of HCMV antigenemia in whole blood. Thus, the development of "in house" techniques can minimize the high costs using commercial kits. Objectives: To optimize and to use Real Time PCR technique "in house" with the Taqman technology for the detection and quantification of betaherpesvírus DNA from HSCT patients. Patients and methods: The primers design was performed of conserved and specific regions for viruses. It was also performed a nested PCR and a comparison between these techniques. Plasma samples from 60 HSCT patients were prospectively obtained, weekly, from the transplantation day until day +100 post-transplant. The results were compared to a control group matched for sex and age and solid organ transplant patients, in this case, heart transplant patients. Results: Of the 60 patients studied, 30 were male and 30 were female. Active HCMV, HHV-6 and HHV-7 infection were detected by nested PCR in plasma in 61.7%, 23.3% and 51.7%, with a median of 34 days (range 2-91 days), 24 days (range 1-96 days) and 15 days (range 0-98 days), respectively. HCMV antigenemia test was positive in 48.3%, in a median range of 40 days (20-91 days) after the transplant. The qPCR optimized for HCMV, HHV-6 and HHV-7 were positive, respectively, in 70%, 16.7% and 41.7%, with a median of 26, 20.5 and 20 days after transplant. Five out of 37 (13.5%) patients with active HCMV infection died in less than 100 days after transplantation (median 50 days, range 46-100 days) and in 2/5 (40%) the main cause of death was HCMV disease. Fourteen patients with active HCMV infection (24.6%) had acute GVHD and overlap. In relation of the number of positive cells for antigenemia, the median was 31 cells. Five patients (8.8%) had disease by HCMV in the gastrointestinal (GI) tract, proven by biopsy. Active HHV-6 infection in plasma was detected in 12/57 patients (21%), in a median of 24 days after transplantation (range 0-84 days). One patient with HHV-6 positive (8.3%) died in less than 100 days after the transplant and had coinfection by HCMV, and died by this agent. Four patients with HHV-6 (33.3%) had acute GVHD and overlap. HHV-7 occurred in 31/57 (54.3%) in a median of 15 days after transplantation (range 0-98 days). Three patients with HHV-7 (9.7%) died in less than 100 days after transplantation and their main cause of death were acute GVHD and bacterial infection. Eight patients with HHV-7 (25.8%) had acute GVHD and overlap. Conclusion: The rapid and early diagnosis of these infections can assist in diagnosis routine and patients treatment that were the focus of this study and other patients who can benefit from our work (AU) | |
| FAPESP's process: | 12/21851-6 - Standardization and Implementation of the technique of Polymerase Chain Reaction in monitoring of early betaherpesvírus reactivation and preemptive therapy in transplant patients of hematopoietic progenitor cells |
| Grantee: | Renato Souza de Oliveira |
| Support Opportunities: | Scholarships in Brazil - Master |