PIX5 Evaluation of diagnostic methods of cytomegalovirus infection in Egyptian renal transplant recipients (original) (raw)
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Background: This study was carried out to detect human cytomegalovirus (HCMV) IgG and IgM antibodies using an Enzyme-linked immunosorbent assay (ELISA) in renal transplant patients in Khartoum state, Sudan and to improve the diagnosis of HCMV through the introduction of Real-time Polymerase Chain Reaction (PCR) testing. A total of 98 plasma samples were collected randomly from renal transplant patients at Ibin Sina Hospital and Salma Centre for Transplantation and Haemodialysis during the period from August to September 2006. Results: Among the 98 renal transplant patients, 65 were males and 33 females. The results revealed that HCMV IgG was present in all patients' plasma 98/98 (100%), while only 6/98 (6.1%) had IgM antibodies in their plasma. HCMV DNA viral loads were detected in 32 patients 32/98 (32.7%) using Real-time PCR.
Cytomegalovirus infection in renal transplant recipients diagnosed by nested-PCR
Brazilian Journal of Medical and Biological Research, 2001
A prospective study of cytomegalovirus (CMV) infection was carried out on 34 renal transplant recipients managed at a General Hospital in Ribeirão Preto, SP, Brazil. Serologic tests showed that all patients were infected with CMV before renal transplantation. Two nested-PCR techniques with primers that recognize sequences of the glycoprotein B (gB) and H (gH) genes were used for CMV detection in blood and urine samples during the post-transplantation period. CMV was detected more frequently in blood samples than in urine samples (P<0.001). Thirty-three patients had CMV detected at least once in blood and/or urine samples. Seven of these patients (21.2%) were diagnosed as having symptomatic CMV infection and showed a worse clinical outcome, with a higher death rate (P = 0.03). No association between CMV viremia and graft rejection was observed. Nested-PCR was not useful to identify patients at risk for symptomatic CMV infection since only 21.2% of the patients with CMV infection were symptomatic.
Cytomegalovirus infection in renal transplant patients
2008
A prospective study of cytomegalovirus (CMV) infection was carried out on 34 renal transplant recipients managed at a General Hospital in Ribeirão Preto, SP, Brazil. Serologic tests showed that all patients were infected with CMV before renal transplantation. Two nested-PCR techniques with primers that recognize sequences of the glycoprotein B (gB) and H (gH) genes were used for CMV detection in blood and urine samples during the post-transplantation period. CMV was detected more frequently in blood samples than in urine samples (P<0.001). Thirty-three patients had CMV detected at least once in blood and/or urine samples. Seven of these patients (21.2%) were diagnosed as having symptomatic CMV infection and showed a worse clinical outcome, with a higher death rate (P = 0.03). No association between CMV viremia and graft rejection was observed. Nested-PCR was not useful to identify patients at risk for symptomatic CMV infection since only 21.2% of the patients with CMV infection were symptomatic.
Incidence and risk factors for cytomegalovirus in kidney transplant patients in Babol, northern Iran
Caspian Journal of Internal Medicine, 2017
Background: Cytomegalovirus (CMV) disease is an important cause of death and possibly transplant rejection in kidney transplant (KT) patients. This study was conducted to investigate the incidence and risk factors of CMV disease in kidney transplant patients. Methods: All end-stage renal disease (ESRD) patients who underwent kidney transplantation during 1998-2014 and their donors were assessed. All samples were followed-up for approximately 70 months. CMV was identified by polymerase chain reaction (PCR) and/or PP65 antigen in peripheral blood leukocytes along with clinical manifestations. Results: A total of 1450 cases participated in the current study. CMV was diagnosed in 178 out of 725 (24.6%) kidney recipients. The annual incidence of CMV disease was 4.2%. Patients older than 40 years had a higher incidence of CMV disease. The level of CMV disease incidence in the 41-60 age group was 4 fold compared to those under 20 of age group (P=0.001). Conclusion: This study demonstrated that the incidence of CMV disease in our region is relatively low and also age more than 40 years and EBV infection are the important risk factors in kidney transplant patients. So care and monitoring of these patients are crucial in the first 5 months.
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.
Medical Principles and Practice, 2007
Objectives: To establish a sensitive and specific real-time PCR for quantitation of cytomegalovirus (CMV) DNA in clinical specimens.Subjects and Methods: In a prospective study, CMV DNA was quantified in blood samples of 255 kidney recipients with and without CMV-related symptoms between the years 2000 and 2005 in Kuwait. In a selected group of patients, the effect of anti-CMV chemotherapy was monitored by quantitative real-time PCR (qRT-PCR). Results: The established qRT-PCR assay had a sensitivity to detect 30 CMV DNA copies. CMV DNA was detected in 54/255 (24%) patients; of these, 17 (31.5%) were asymptomatic, and 37 patients (68.5%) had symptomatic CMV infection. Sequential blood specimens were collected from all CMV-positive patients and tested by CMV pp65 antigenemia and qRT-PCR assays. There was a moderate positive correlation between the two assays (Pearson’s correlation = 0.52). The median CMV viral load measured by qRT-PCR was higher in symptomatic (6.5 × 104 copies/ml) th...
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.
Transplant International, 1998
Detection of human cytomegalovirus (CMV) at an early stage of infection is a prerequisite for the initiation of effective antiviral therapy. Nucleic acid sequence-based amplification (NASBA) has been designed for the specific amplification of RNA. NASBA proved to be a highly sensitive method for the detection of human immunodeficiency virus mRNA . The technique has now also been applied to the qualitative detection of CMV. Two different viral mRNAs were chosen as targets for amplification by NASBA, the immediate early (IE) 1 mRNA (UL123) [7] and the late pp67 mRNA (UL65) . These mRNAs are synthesized at different phases in the replication cycle of CMV. IE mRNA is transcribed within a few hours after infection of the cell, while the late phase of transcription is initiated approximately 72 h after infection and requires the active replication of the CMV genome. Blood specimens from patients with a kidney allograft were screened for the presence of IE and late mRNA. Results were compared with virus isolation, pp65 antigenemia and serology, in order to evaluate the diagnostic value of the IE and late NASBA assays.
Use of Laboratory Assays To Predict Cytomegalovirus Disease in Renal Transplant Recipients
Journal of Clinical Microbiology, 1998
Eight laboratory assays, viz., the pp65 direct antigenemia test, a quantitative cytomegalovirus (CMV)-specific immunoglobulin G (IgG) assay (Biomerieux VIDAS), a CMV-specific IgM assay (Biomerieux VIDAS), the Hybrid Capture system (Murex), an in-house PCR with plasma (P-PCR) and leukocytes (L-PCR), and a commercial PCR (Roche AMPLICOR) with plasma (P-AMP) and leukocytes (L-AMP), were compared for their abilities to predict CMV disease before the onset of illness in a prospective study of 37 renal transplant recipients. By using an expanded criterion for active infection (two or more of the markers positive) and a clinical definition of disease, 22 (59%) patients were identified as having active CMV infection and 13 (35%) were identified as having CMV disease. Of the 13 CMV-seronegative recipients who received seropositive kidneys (R− group), 8 had active infection and disease. All assays were 100% specific and 100% predictive of CMV disease in the R− group. The leukocyte PCRs (L-PCR...