Comparison of ELISA, Antigenemia Assay and Nested PCR Monitoring Techniques for Detection of Cytomegalovirus Infection in Renal Transplantation Patients

Ehsan Moghanloo,Vahid Babaei,Hamidreza Zeraatgar Gohardani,Tina Delsouz Bahri,Soheila Rezaei,Leila Khanmirzaei,Shahram Teimourian

doi:10.4172/2161-0703.1000274

Abstract

Human cytomegalovirus (HCMV) can be transmitted through blood transfusion and organ transplantation and could be cause of some complication in solid-organ transplant recipients. Current study is aimed to compare the sensitivity and Specificity of ELISA, Antigenemia assay and nested PCR methods to detection of Cytomegalovirus infection in renal transplantation patients. In this study blood samples were collected from 200 renal transplant recipients’ patients. DNA was extracted by commercial kit and Nested PCR was done by 2 pairs of internal and external primers. Anti CMV antibodies (IgM and IgG) were detected by ELISA and CMV-pp65 antigenemia assay (Ag) was used to detect CMV antigens. The sensitivity and Specificity of each test and all the methods together were evaluated, and SPSS software was used to analysis of data. From 200 patients, 193 (96.5%) were positive for CMV antibodies with the Specificity of 100 and sensitivity of 97.76%. 120 (60%) and 25 (12.5) samples were positive by nested PCR and Ag assay with the Specificity of 94.49 and 78.12 and sensitivity of 94.49 and 78.12, respectively. In the case of early diagnosis of the disease, nested PCR diagnose the infection 14 years earlier than Ag assay and was consistently positive, whereas false negative results were frequently observed with the pp65 Ag assay. The sensitivity and specificity of the two methods combined detection for CMV infection were 96.76% and 99.89%. ELISA can be used as a screening reliable detection test for CMV infection in recipient especially when PCR is unavailable. Combination of ELISA and CMV-PCR methods, provide a more effective method to monitor CMV infection.

Highlights

Cytomegalovirus (CMV) is one of the most important causes of complication in solid-organ transplant recipients and this infection has a significant effect on transplant outcome and is common in renal transplant recipients [1,2]
There was no relation between CMV infection, age and gender of the patients (P=0.183)
Results of different CMV infection monitoring methods A total of 200 recipients were analysed by nested PCR, Anti-CMV

Summary

Introduction

Cytomegalovirus (CMV) is one of the most important causes of complication in solid-organ transplant recipients and this infection has a significant effect on transplant outcome and is common in renal transplant recipients [1,2]. There are no specific symptoms of CMV infection after renal transplantation in patients, but this infection causes many diverse clinical outcomes. It is difficult to accurately diagnose CMV infection. It is essential to use different laboratory diagnostic methods to diagnose CMV infection [3,4]. The availability of a sensitive test capable of detecting CMV at an early stage of infection is so essential. The antigenemia assay (Ag) is a reference method commonly used for CMV virus quantification in blood specimens. Pp antigen (pp Ag) is measured by the quantitation of positive leukocyte nuclei and has been validated in clinical trials with immunosuppressed patients [5,6] The antigenemia assay (Ag) is a reference method commonly used for CMV virus quantification in blood specimens. pp antigen (pp Ag) is measured by the quantitation of positive leukocyte nuclei and has been validated in clinical trials with immunosuppressed patients [5,6]

Methods

Results

Discussion

Conclusion