Microbiologia Médica e Diagnóstico

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

Abstract

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

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.