Monitoring of Cytomegalovirus Reactivation in Bone Marrow Transplant Recipients by Real-time PCR

2008 ◽  
Vol 14 (4) ◽  
pp. 399-409 ◽  
Author(s):  
Seyed H. Ghaffari ◽  
Narghes Obeidi ◽  
Mehdi Dehghan ◽  
Kamran Alimoghaddam ◽  
Ahmad Gharehbaghian ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Real Time Pcr ◽  
Marrow Transplant ◽  
Transplant Recipients ◽  
Cytomegalovirus Reactivation

scholarly journals Monitoring of Active HHV-6 Infection in Bone Marrow Transplant Recipients by Real Time PCR; Comparison to Detection of Viral DNA in Plasma by Qualitative PCR

2002 ◽  
Vol 46 (10) ◽  
pp. 701-705 ◽  
Author(s):  
Masaru Ihira ◽  
Tetsushi Yoshikawa ◽  
Kyoko Suzuki ◽  
Masahiro Ohashi ◽  
Sadao Suga ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Real Time Pcr ◽  
Marrow Transplant ◽  
Transplant Recipients ◽  
Viral Dna ◽  
Qualitative Pcr

scholarly journals Quantification of Human Cytomegalovirus DNA in Bone Marrow Transplant Recipients by Real-Time PCR

2001 ◽  
Vol 39 (12) ◽  
pp. 4362-4369 ◽  
Author(s):  
F. Griscelli ◽  
M. Barrois ◽  
S. Chauvin ◽  
S. Lastere ◽  
D. Bellet ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Human Cytomegalovirus ◽  
Real Time Pcr ◽  
Marrow Transplant ◽  
Transplant Recipients

scholarly journals Detection of invasive aspergillosis in bone marrow transplant recipients using real-time PCR

2013 ◽  
Vol 5 (2) ◽  
pp. 68 ◽  
Author(s):  
Tahereh Shokohi ◽  
Ghasem Janbabaie ◽  
MohammadBagher Hashemi-Soteh ◽  
Kamran Ali-Moghaddam ◽  
SeyedReza Aghili ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Invasive Aspergillosis ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Real Time Pcr ◽  
Marrow Transplant ◽  
Transplant Recipients

scholarly journals Detection of Human Polyomaviruses in Urine from Bone Marrow Transplant Patients: Comparison of Electron Microscopy with PCR

2004 ◽  
Vol 50 (2) ◽  
pp. 306-312 ◽  
Author(s):  
Stefan S Biel ◽  
Andreas Nitsche ◽  
Andreas Kurth ◽  
Wolfgang Siegert ◽  
Muhsin Özel ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Bone Marrow ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Real Time Pcr ◽  
Marrow Transplant ◽  
Test System ◽  
Urine Samples ◽  
Clinical Samples ◽  
Transplant Patients

Abstract Background: We studied electron microscopy (EM) as an appropriate test system for the detection of polyomavirus in urine samples from bone marrow transplant patients. Methods: We evaluated direct EM, ultracentrifugation (UC) before EM, and solid-phase immuno-EM (SPIEM). The diagnostic accuracy of EM was studied by comparison with a real-time PCR assay on 531 clinical samples. Results: The detection rate of EM was increased by UC and SPIEM. On 531 clinical urine samples, the diagnostic sensitivity of EM was 47% (70 of 149) with a specificity of 100%. We observed a linear relationship between viral genome concentration and the proportion of urine samples positive by EM, with a 50% probability for a positive EM result for urine samples with a polyomavirus concentration of 106 genome-equivalents (GE)/mL; the probability of a positive EM result was 0% for urine samples with <103 GE/mL and 100% for urine samples containing 109 GE/mL. Conclusions: UC/EM is rapid and highly specific for polyomavirus in urine. Unlike real-time PCR, EM has low sensitivity and cannot quantify the viral load.

scholarly journals Detection of Aspergillus species in bone marrow transplant patients

2010 ◽  
Vol 4 (08) ◽  
pp. 511-516 ◽  
Author(s):  
Parisa Badiee ◽  
Abdolvahab Alborzi
Keyword(s):  
Bone Marrow ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Real Time Pcr ◽  
Clinical Signs ◽  
Marrow Transplant ◽  
Aspergillus Species ◽  
Pcr Assay ◽  
Blood Samples ◽  
Transplant Patients

Introduction:  Invasive aspergillosis is a severe complication of cytotoxic chemotherapies and bone marrow transplantation (BMT). The aim of this study was to assess the utility of a real-time PCR assay for the early diagnosis of Aspergillus species in blood samples from BMT patients. Methodology: Blood specimens (n = 993) from patients (n = 82) scheduled for BMT were collected prior to transplant and for 100 days post transplantation.  The specimens were later tested using an Aspergillus-specific real-time PCR assay. Cultures of clinical samples, along with sonography and computerized tomographic scans, were performed as standard of care. Results: Aspergillus DNA was positive in 94 sequential blood samples from 13 patients with clinical and radiological signs of infection. Samples from three of these patients were PCR-positive for Aspergillus in the first week of admission, prior to transplantation. Four patients with aspergillosis were cured with antifungal agents and nine died. An additional 12 patients without clinical signs of infection were PCR-positive on one occasion each, while two patients with clinical signs of infection were PCR-negative. Compared to routine methods of aspergillosis diagnosis, the respective sensitivity, specificity, negative, and positive predictive values of the PCR method by patient were 86.6%, 82%, 96.5% and 52%. Conclusions: The results show that Aspergillus infections in the blood of bone marrow transplant patients can be dectected by PCR methods. Early detection of Aspergillus infections by PCR has the potential to positively impact patient mortality rate and provide cost savings to hospitals.

scholarly journals Quantitative Real-Time PCR with Automated Sample Preparation for Diagnosis and Monitoring of Cytomegalovirus Infection in Bone Marrow Transplant Patients

2004 ◽  
Vol 50 (5) ◽  
pp. 846-856 ◽  
Author(s):  
Kyeong Man Hong ◽  
Hazim Najjar ◽  
Mary Hawley ◽  
Richard D Press
Keyword(s):  
Bone Marrow ◽  
Viral Load ◽  
Antiviral Therapy ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Real Time Pcr ◽  
Marrow Transplant ◽  
Diagnostic Sensitivity ◽  
Pcr Assay ◽  
Transplant Patients

Abstract Background: In bone marrow and stem cell transplant patients, the widespread use of preemptive cytomegalovirus (CMV) antiviral therapy necessitates faster, more precise, and more sensitive quantitative laboratory methods for serial viral load monitoring. Methods: We developed a novel CMV viral load assay using real-time PCR of plasma DNA prepared by an automated robotic workstation. Fluorescent hybridization probes directed at the glycoprotein B (gB) gene (or EcoRI D region) of CMV were used to detect and quantify PCR products. The β-globin gene was amplified in parallel to control for the efficiency of the extraction and PCR steps. Results: The assay was linear (R = 0.999) from a lower detection limit of 125 copies/mL to 5 × 109 copies/mL with a PCR efficiency of 1.975 (gB) or 2.02 (EcoRI D). The viral loads determined by PCRs directed at these two different viral targets were no different (n = 53; R = 0.928). The interassay CV was 3.5%, and the intraassay CV was 1–4%. Compared with a commercially available quantitative competitive PCR assay (Roche MONITOR; R = 0.59), the mean CMV viral load by real-time PCR was 3.1 times higher (mean ratio; P = 0.002). The diagnostic sensitivity and specificity of the real-time assay were 96% and 100%, respectively (n = 147), compared with 74% and 98% for a qualitative PCR assay (Roche AMPLICOR). On a subset of samples, the diagnostic sensitivity of viral culture was no greater than 50% (n = 44). Of 1115 clinical referral samples from 252 patients, 10% of the samples and 18% of the patients had low-level CMV viremia (median, 500 copies/mL). In this predominantly (85%) bone marrow transplant testing cohort, serial CMV viral load results were the predominant clinical trigger for the initiation, monitoring, and cessation of preemptive antiviral therapy. Conclusions: The combination of automated DNA preparation and semiautomated real-time fluorescent PCR detection allows for a sensitive, precise, and accurate high-throughput assay of CMV viral load that can be used as the laboratory trigger for preemptive antiviral therapy.

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