Comparison of Real-time PCR to ELISA for the detection of human cytomegalovirus infection in renal transplant patients in the Sudan (original) (raw)
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Sudan Medical Monitor, 2013
Introduction: Human cytomegalovirus (HCMV) is a member of the genus Herpes virus and belongs to the family Herpesviridae. Objective: The aim of this research work was to study the prevalence of human cytomegalovirus (HCMV) infection in renal transplant and haemodialysis patients. Blood samples were collected randomly from 52 renal transplant patients and 41 haemodialysis patients. The sera were tested with an enzyme linked immunosorbent assay (ELISA) for HCMV IgG antibodies and additional ELISA test for HCMV IgM antibodies. Results: Renal transplant screening revealed that 98% of patients have IgG for HCMV antibodies and only 6% have IgM antibodies. In haemodialysis patients 95% showed the presence of IgG antibodies to HCMV and non of patient revealed the presence of IgM antibodies.
Detection of cytomegalovirus using PCR in serum from renal transplant recipients
Journal of Clinical Pathology, 1995
Aims-To develop a polymerase chain reaction (PCR) assay for the detection of cytomegalovirus (CMV) DNA in serum and leucocytes of renal transplant recipients and compare this assay with CMV culture and serodiagnosis. Methods-Monthly specimens were obtained from 12 patients starting immediately before transplant. CMV infection was monitored by IgM enzyme linked immunosorbent assay, virus culture and PCR on serum and leucocytes. Results-Two offour IgG positive patients had reactivation of CMV disease confirmed by culture, three of eight seronegative patients had a primary infection, one confirmed by serology and two by culture. PCR was positive earlier than conventional methods in three cases and concurrently in two. No positive PCR reactions occurred in the seven patients who remained negative by culture and serology. Conclusions-CMV DNA is detectable in serum; serum may be positive before virus is detectable by buffy coat culture; and PCR may be useful as an early indication of potential CMV disease in renal transplant recipients.
2010
Objective: Cytomegalovirus (CMV) has been recognized as one of the most important opportunistic pathogens in kidney transplant patients. The aim of this study was to determine the prevalence of CMV antibody in donors and recipients before transplantation. Material and Methods: In a cross sectional study from March 2008 to August 2009 we prospectively studied donors and recipients who referred to our kidney transplant center. All of routine pretransplante laboratory studies including liver function tests and CMV IgG and IgM antibody were performed for them. Results: A total of 148 patients (79 donors and 69 recipients) were included in the study. Mean age of donors and recipients were 30 ± 8 years and 40 ± 18 years respectively. Liver Function tests (SGOT and SGPT) were at normal range and marker of HBV infection was negative in both groups but HCV antibody was positive in 2.89 percent of recipients (n=2) and negative in all of donors. CMV IgG antibody was positive in 100 percent of ...
Transplantation Proceedings, 2005
Cytomegalovirus infection is a common complication of renal transplantation. Antigen pp65 levels serve as indicators of viral load, although the technique is difficult to perform and interpret. We sought to determine whether quantitative PCR had a higher sensitivity and predictive value in CMV infection. Methods. The study included 100 renal transplant recipients who were screened for IgM and IgG at the time of admission. On days 7, 30, 45, 60, 75, 90, 120, 180, and 360, antigenemia tests were performed on blood (pp65) and urine, and a quantitative PCR on blood. Among 59 patients recruited between November 2003 and August 2004 the mean age was 54.5 Ϯ 12.9 years. Two patients did not reach 90 days follow-up (3%); four patients have not surpassed 90 days (7%); 22, 120 days (37%); and 31, 180 days (53%). Ninety-three percent of patients showed anti-IgG CMV-positive titers with all being IgM CMV-negative at baseline. The patients at risk for infection were given valgancyclovir as prophylaxis throughout the study. Results. At 474 visits, 8 samples (2.4%) were positive with urine; 5 (1.4%) with pp65, and 15 (4.7%) with PCR. Among the 15 positive samples, two (Ͼ100,000 and 3250 copies) revealed agreement of positive IgM and shellvial test on urine; two (15,100 and 5670 copies), antigen pp65 1ϩ; one (17,400 copies) with pp65 2ϩ and shellvial urine; two (99,400 and 28,300 copies) with pp65 1ϩ and shellvial urine; and eight remaining determinations, 749, 2250, 686, 928, 2250, 26600, 777, and 2790 copies. The rest of the tests were negative. Conclusion. The preliminary results of this study demonstrated that quantitative PCR was a useful rapid tool for diagnosing and monitoring CMV infections.
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.
International Journal of Infectious Diseases, 2002
Background: Quantitative monitoring of human cytomegalovirus (HCMV) is currently used in the follow-up of immunosuppressed patients. Objective: To investigate whether real-time PCR quantification (QPCR) of HCMV DNA could replace pp65 antigenemia. Study design: We compared HCMV QPCR on whole blood (WB) and on plasma with a pp65-antigenemia assay on 192 samples. Afterwards, we tested 1310 samples from 308 immunosuppressed patients both by antigenemia assay and QPCR on WB. Results: The first study comparison showed that QPCR results on WB and plasma were significantly correlated with antigenemia. QPCR on WB was more sensitive than QPCR on plasma or antigenemia, detecting 31 and 49 additional positive samples, respectively. During the second comparison, QPCR on WB and antigenemia were again correlated (r = 0.70; p < 0.0001), but QPCR detected 244 additional positive samples. HCMV DNA was detected earlier than pp65 antigen (median difference: 14 days; range: 7-30). One, 5, 10, 50 and 100 pp65-positive cells/200,000 leukocytes corresponded to 439, 1531, 2623, 9150 and 15,671 HCMV DNA copies/mL of WB, respectively, but this equivalence differed according to the sub-group of patients considered. Conclusion: QPCR on WB is the most sensitive method for the monitoring of HCMV infection in immunosuppressed patients.
Transplantation Proceedings, 2007
Human cytomegalovirus (CMV) infection is a major cause of morbidity and mortality among kidney transplant recipients. The CMVpp65 antigenemia assay has been used for preemptive therapy. Real-time polymerase chain reaction (PCR) technology for CMV DNA quantification in blood has demonstrated a good correlation with the currently employed CMV antigenemia assay. In this study, 90 renal transplant recipients were prospectively enrolled from July 2004 and May 2005. Monitoring of CMV infection was routinely performed with CMV antigenemia and real-time PCR assays. Real-time plasma PCR and CMV antigenemia assays were assessed on 797 samples. CMV antigenemia correlated with a positive CMV PCR (2 ϭ 78.05; P Ͻ .0001). Not only the positive rate but also the number of positive cells correlated with the number of PCR DNA copies (F ϭ 26.07, r 2 ϭ .25, P Ͻ .0001). To define an optimal cutoff value of CMV DNA load to initiate treatment in kidney transplant patients, we considered a CMV antigenemia titer of Ͼ50 positive cells per 400,000 leukocytes as the gold standard in our previous study. The optimal cutoff value for the quantitative real-time PCR assay was predicted to be 86 copies/L. Thus, we observed that CMV real-time PCR assay would not completely replace antigenemia assay in kidney transplant recipients, but can be used complementarily to screen antigenemia and monitor preemptive therapy.