Significance of nested PCR and quantitative real time PCR for cytomegalovirus detection in renal transplant recipients

Kanaan, Atef Kanaan; Cour, I. La; Álvarez‐Lafuente, Roberto; M, Benedicto; Culebras, Esther; Prats, D; Fernández, Cristina; Picazo, Juan J.

doi:10.1016/j.ijantimicag.2004.06.012

Cited by 7 publications

(6 citation statements)

References 28 publications

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“…26 -30 In spite of the obtained statistical association, not all the samples resulted positives by both PCR methods, especially those from pregnant women with non primary active infection, which could be explained by the fact that the sensitivity of the quantitative method is higher, which had been previously reported in the literature. 31,32 More recently, the availability of quantitative PCR-assays has increased interest in the potential role of viral load in predicting disease outcome, development of hearing loss and monitoring response to antiviral therapy in congenital HCMV infection; however, in practice many questions are still open. 2 Revello et al, and Romanelli et al, have not found association between the viral load and the prediction of the symptom in the newborns or the appearance of sequelae 33,34 which is in agreement with the lack of associations also reported in the present study.…”

Section: Discussionmentioning

confidence: 99%

Diagnosis and Screening for Cytomegalovirus Infection in Pregnant Women in Cuba as Prognostic Markers of Congenital Infection in Newborns

Kourí¹,

Correa²,

Verdasquera³

et al. 2010

Pediatric Infectious Disease Journal

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Section: Discussionmentioning

confidence: 99%

Diagnosis and Screening for Cytomegalovirus Infection in Pregnant Women in Cuba as Prognostic Markers of Congenital Infection in Newborns

Kourí¹,

Correa²,

Verdasquera³

et al. 2010

Pediatric Infectious Disease Journal

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“…It is considered that positive DNA in serum/plasma is connected more closely with symptomatic infection [ 15 , 16 ], because serum/plasma DNA reflects disruption of host cells caused by active viral replication. In other words, it seems that PCR using PBL DNA is not suitable for diagnosing active CMV infection, because healthy adults with the latent infection may harbor CMV in their leukocytes.…”

Section: Discussionmentioning

confidence: 99%

“…Some reports [ 12 - 14 ] suggest that leukocyte-based tests are superior for detecting HCMV DNA. On the other hand, several studies show that plasma/serum positivity is more correlated with active infection and the processing of plasma is easier [ 15 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

Monitoring human cytomegalovirus infection with nested PCR: comparison of positive rates in plasma and leukocytes and with quantitative PCR

Shu

Zhou

et al. 2010

Virol J

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BackgroundHuman cytomegalovirus (HCMV) infection poses a significant health threat to immunocompromised individuals. Here we performed this study to set up a highly sensitive nested PCR method applicable for detecting HCMV infection in high-risk individuals. In this work, 106 blood specimens from 66 patients with potential HCMV infection were obtained. Total DNA was extracted separately from plasma and peripheral blood leukocytes (PBL) of each sample. HCMV DNA was detected in parallel by nested PCR and quantitative real-time PCR (qRT-PCR), and the results were compared.ResultsSerial dilution test revealed that the detection limit of nested PCR was 180 copies/ml. The nested PCR showed a higher positive rate than qRT-PCR (34.9% vs. 12.3%, p < 0.001). The positive rate of nested PCR based on PBL DNA was significantly higher than that based on plasma DNA (34.9% vs. 18.9%, p = 0.002). Of the 14 patients with serial samples, 11 were positive for HCMV DNA in PBL while only 7 were positive in plasma. Moreover, for each patient, nested PCR using PBL DNA also detected more positive samples than that using plasma DNA.ConclusionCombined use of nested PCR with PBL DNA is highly sensitive in defining HCMV infection. This assay is particularly useful in the case of quantification not essential.

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“…Briefly, the TTV genomic DNA was extracted from blood using a dinitrophenol kit (Cinna Gen Inc., Tehran, Iran) according to manufacturer instruction. The presentation of TTV genomic DNA was analyzed in ALL patients using an in-house semi-nested-PCR protocol, as previously described [ 35 , 38 , 62 ].…”

Section: Methodsmentioning

confidence: 99%

MiR-181a and -b expression in acute lymphoblastic leukemia and its correlation with acute graft-versus-host disease after hematopoietic stem cell transplantation, COVID-19 and torque teno viruses

et al. 2021

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Acute lymphoblastic leukemia (ALL), a malignant transformation and proliferation of the lymphoid line of blood cells, is characterized by chromosomal abnormalities and genetic changes. The purpose of this research was the evaluation of expression level of miR-181a and -b in patients with ALL compared to the control group. Furthermore, we examined their expression level in hematopoietic stem-cell transplantation (HSCT) patients who developed acute graft-versus-host disease (aGVHD) in comparison with those without aGVHD and explore the relationship between their expression level and cytogenetic abnormalities. In this cross-sectional study, 76 newly diagnosed adult De novo ALL patients were enrolled who were admitted to our referral hospital. All patients received standard chemotherapy, consisting of daunorubicin. A total of 37 patients underwent HSCT from the related human leukocyte antigen-matched donors. ALL patients have been diagnosed with the coronavirus disease 2019 (COVID-19) and Torque teno viruses (TTVs). We assessed the expression levels of miR-181a and -b in the peripheral blood sample of ALL patients at the time of diagnosis prior to chemotherapy, and healthy matched individuals by RT–PCR. TTVs and COVID-19 load were also determined via RT–PCR. In conclusion, the expression level of miR-181a and -b were significantly higher in ALL patients than healthy controls and also increased in patients who developed aGVHD in comparison with those without aGVHD. MiR-181a and -b can be a useful biomarker in ALL and a useful indicator of aGVHD. The expression level of miR-181a in ALL patients with COVID-19 is significantly up-regulated, while it is reduced in these patients with TTV.

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Significance of nested PCR and quantitative real time PCR for cytomegalovirus detection in renal transplant recipients

Cited by 7 publications

References 28 publications

Diagnosis and Screening for Cytomegalovirus Infection in Pregnant Women in Cuba as Prognostic Markers of Congenital Infection in Newborns

Diagnosis and Screening for Cytomegalovirus Infection in Pregnant Women in Cuba as Prognostic Markers of Congenital Infection in Newborns

Monitoring human cytomegalovirus infection with nested PCR: comparison of positive rates in plasma and leukocytes and with quantitative PCR

MiR-181a and -b expression in acute lymphoblastic leukemia and its correlation with acute graft-versus-host disease after hematopoietic stem cell transplantation, COVID-19 and torque teno viruses

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