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      "pp65 antigenemia and real time polymerase chain reaction (PCR) based-study to determine the prevalence of human cytomegalovirus (HCMV) in kidney donors and recipients with follow-up studies."

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          Abstract

          Background

          The present study was undertaken to determine the rate of occurrence of Human cytomegalovirus (HCMV) among kidney transplant recipients and donors by application of direct detection methods and to understand HCMV infection/disease development among transplanted patients as a prospective study.

          Results

          Peripheral blood samples collected from 76 kidney donors and 76 recipients from September 2007 to August 2009 were subjected to pp65 antigenemia and Quantitative real-time PCR (qRT-PCR) assays. Data were analyzed under Group A, B and C. Group A was further divided into sub-groups I, II, III, IV, and V for better understanding. Three, one and two donors in sub-group I, III, IV of Group A tested positive for real time PCR respectively. One recipient from group III tested positive for HCMV by qRT- PCR prior transplantation and remained positive one month post-transplantation. Three other recipients, tested negative prior to transplantation became positive a month after transplantation. Group B consisted of 18 donor-recipient pairs and one of the donor tested positive for HCMV by qRT-PCR. Eight recipients tested positive for HCMV one month after transplantation. The pp65 positivity and HCMV DNA load was high among group C recipients who mostly had symptoms of active disease. Significantly high values of pp65 antigenemia were observed among recipients of sub-group II (non-parametric chi-square test p = 0.007). Positive correlation between pp65 antigenemia and qRT-PCR value was observed. Thirty three of the recipients with disease treated with Valgancyclovir showed improved clinical outcome.

          Conclusion

          Our study showed that a significant proportion of kidney recipients develop HCMV infection following renal transplantation in spite of the absence of HCMV among donors. pp65 antigenemia assay and qRT- PCR methods can be applied to detect HCMV among kidney donors and recipients to monitor development of disease and these assays were predicative of HCMV infection among them. Clinical resistant to valganciclovir was not observed.

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          Most cited references17

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          Seroprevalence of cytomegalovirus among voluntary blood donors in Delhi, India.

          Cytomegalovirus (CMV) is known to be a significant cause of morbidity and mortality following blood transfusion in children and immunocompromised adults. In India, it is not mandatory to screen donated blood for CMV in blood banks. Very few studies have been conducted in India to estimate the seroprevalence of this infection in voluntary blood donors. This study was conducted to estimate the seroprevalence of CMV among voluntary blood donors in Delhi, India. In this study, none of 200 donors tested positive for CMV IgM antibody, but 95% were positive for CMV IgG antibody. There was no statistically significant difference in seropositivity of CMV based on distribution of age. Of the 200 donors, 3% tested positive for HBsAg, 1% for HIV, 2% for hepatitis C virus, and 4.5% for syphilis. Since about 95% of blood donors in India are seropositive for CMV, it would seem superfluous to screen blood donors for CMV, as very few seronegative blood units would be available for transfusion. Other preventive strategies, such as leukoreduction, etc., could be more appropriate and cost-effective for the prevention of transmission of CMV through infected blood to immunosuppressed individuals.
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            Monitoring human cytomegalovirus infection in transplant recipients.

            Human cytomegalovirus (HCMV) infection remains one of the most challenging infectious complications in both solid organ transplant (SOT) and hemopoietic stem cell transplant (HSCT) recipients. In the last two decades advances have been made in the diagnosis and monitoring of HCMV infection in SOT and HSCT recipients following introduction of quantitative assays such as rapid virus isolation in blood (viremia), quantitation of pp65 in peripheral blood leukocytes (antigenemia), and quantitation of viral genome in blood (DNAemia). The availability of these rapid diagnostic assays has allowed treatment administration during the presymptomatic phase of HCMV infection (preemptive treatment) and greatly reduced HCMV-related morbidity and mortality, particularly among HSCT recipients. Definition of clinically validated thresholds for initiating preemptive treatment in SOT and HSCT recipients is a major goal in the transplantation setting. With respect to universal prophylaxis of HCMV infection in transplant recipients, the preemptive treatment approach shows advantages in (i) treating a lower number of patients for shorter periods of time and (ii) avoiding the reported emergence of HCMV disease after interruption of anti-HCMV prophylaxis. To understand the mechanism behind long-term control of HCMV infection in transplant recipients, the HCMV-specific T-cell response must be evaluated.
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              Relationship between pp65 antigenemia levels and real-time quantitative DNA PCR for Human Cytomegalovirus (HCMV) management in immunocompromised patients

              Background Quantitative real-time PCR assays, which are more rapid and practical than pp65 antigenemia determination, are progressively becoming the preferred method for monitoring Human Cytomegalovirus (HCMV) reactivation. However, the relationship between HCMV DNA and antigenemia levels is still under investigation. The aim of this study was to analyse the relationship between HCMV DNA and pp65 antigenemia levels in order to identify clinically useful threshold values for the management of patients. Methods 475 consecutive samples from 156 immunosuppressed patients were tested for HCMV by pp65 antigenemia and Real-time PCR assay. Results 136 out of 475 consecutive samples derived from 48 patients showed evidence of HCMV infection. HCMV DNA was detected in 106 samples, pp65 antigen in 3, and both markers in 27. pp65 antigen detection was associated with higher HCMV DNA levels. The cut-off HCMV DNA level that best predicted pp65 antigenemia in this series of samples was 11,500 copies/ml, but different threshold levels could be observed for specific groups of patients. HCMV disease was observed in 5 out of 48 patients with active HCMV infection. The presence of clinical symptoms was associated with positive pp65 and with higher antigenemia levels. Higher HCMV DNA load at the onset of viral replication was correlated to the development of clinical symptoms. Conclusion Both pp65 antigenemia and HCMV DNA load can be useful for the prospective monitoring of immunocompromised subjects. Specific cut-off levels capable of triggering preemptive antiviral treatment should be determined in accordance to the type of test used and the characteristics of patients and prospectively validated.
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                Author and article information

                Journal
                Virol J
                Virology Journal
                BioMed Central
                1743-422X
                2010
                16 November 2010
                : 7
                : 322
                Affiliations
                [1 ]L & T Micobiology Research Center, Sankara Nethralaya, 18, College Road, Chennai - 600 006. India
                [2 ]Kaliappa Renal Centre, Billroth Hospitals, Chennai, India
                Article
                1743-422X-7-322
                10.1186/1743-422X-7-322
                2998481
                21080922
                f538c147-ff9d-471e-b435-9e020c552e57
                Copyright ©2010 Madhavan et al; licensee BioMed Central Ltd.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 9 August 2010
                : 16 November 2010
                Categories
                Research

                Microbiology & Virology
                Microbiology & Virology

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