Significance of Qualitative Polymerase Chain Reaction Combined with Quantitation of Viral Load in the Diagnosis and Follow-Up of Cytomegalovirus Infection after Solid-Organ Transplantation

Intervirology ◽  
1999 ◽  
Vol 42 (5-6) ◽  
pp. 398-404 ◽  
Author(s):  
H. Vanpoucke ◽  
B. van Vlem ◽  
R. Vanholder ◽  
L. van Renterghem
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Load ◽  
Organ Transplantation ◽  
Cytomegalovirus Infection ◽  
Solid Organ Transplantation ◽  
Solid Organ ◽  
Chain Reaction ◽  
Qualitative Polymerase Chain Reaction ◽  
Polymerase Chain

Quantitation of cytomegalovirus DNA by the polymerase chain reaction as a predictor of disease in solid organ transplantation

2004 ◽  
Vol 73 (2) ◽  
pp. 223-229 ◽  
Author(s):  
Valeria Ghisetti ◽  
Anna Barbui ◽  
Alessandro Franchello ◽  
Silvia Varetto ◽  
Fabrizia Pittaluga ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Organ Transplantation ◽  
Solid Organ Transplantation ◽  
Solid Organ ◽  
Chain Reaction ◽  
Polymerase Chain

Adapted Management of EBV Reactivation After Solid Organ Transplantation: An Effective Prevention of Post Transplantation Lymphoproliferative Disorders (PTLD). Results of the Largest Prospective Study on 251 Patients

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 592-592
Author(s):  
Sylvain Choquet ◽  
Shaida Varnous ◽  
Claire Deback ◽  
Alain Pavie ◽  
Véronique Leblond
Keyword(s):  
Viral Load ◽  
Board Of Directors ◽  
Organ Transplantation ◽  
Solid Organ Transplantation ◽  
First Year ◽  
Solid Organ ◽  
Post Transplantation ◽  
Advisory Committees ◽  
Ebv Reactivation

Abstract Abstract 592 Background: PTLD represent a rare but aggressive graft complication. Patients who have received a solid organ transplantation have a 20 to 120 fold higher incidence of non-Hodgkin's lymphoma. EBV reactivation represents a major predictive factor for PTLD, especially during the first year after transplantation, but there is no consensual attitude in this situation Aim: We conducted a monocentric prospective study in the Hospital of Pitie Salpêtriere, Paris, France, on all new heart transplanted patients. EBV viral load (EVL) on whole blood samples was systematically followed and confirmed reactivations were treated depending on viral load. Methods: All heart transplanted patients who had at least one EVL between January 2004 and December 2008 were included. Immunosuppression consisted on anti-lymphocyte sera, ciclosporin, mycophenolate-mofetyl (MM) and prednisone. Twelve to 15 blood samples per year were analysed. If the EVL was more than 105 copies/ml, a CT scan or a PET-san was performed in order to detect any PTLD and patients were treated by diminution of the immunosupression (DIS), mainly by MM arrest. One injection of Rituximab (R) (375 mg/m2) was used in case of failure and/or if EVL was over 106 copies/ml. Results: A total of 251 patients were included, 59 femals/192 men, of a median age of 50 years [16-72]. All but 6 were EBV positive before the graft. Reactivations were detected in 29 cases (11,55%) and treated by DIS only in 20 cases, DIS followed by R in 5 and directly by DIS and R in 4. All EBV negative patients developed a primoinfection in the first year, 2 with an EVL over 105, one presented non documented hepatic lesions which disappeared after DIS. All EBV reactivations were controlled, with a relapse in only one case (reactivations treated the first time by DIS, 10 months later by DIS and R and 6 months later by DIS). With a median follow-up of 1118 days [53-2100] only one PTLD has been diagnosed (in a patient lost to follow up and taken in charge in an other unit) and 24 patients died (9,5%). Analyse of DIS +/− R on graft rejection and potential link between CMV reactivation and EBV reactivation will be presented at the ASH. From 1987 to December 2003, 24 (1,8/year) PTLD have been treated in the same unit (18 EBV positive, 5 negative, 1 unknown), of which 13 were early PTLD (all EBV positive) diagnosed within one year post transplantation. Conclusions: EBV reactivation after organ transplantation can be managed by diminution of immunosupression and/or rituximab, depending on viral load, without serious complication. This adapted management seems to decrease dramatically the incidence of EBV positive PTLD. Disclosures: Choquet: ROCHE: Consultancy. Leblond:ROCHE: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; MUNDIPHARMA: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; CELGENE: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees.

Nested Polymerase Chain Reaction With Sequence-Specific Primers Typing for HLA-A, -B, and -C Alleles: Detection of Microchimerism in DR-Matched Individuals

Blood ◽  
1999 ◽  
Vol 94 (4) ◽  
pp. 1471-1477 ◽  
Author(s):  
Anthony S. Carter ◽  
Lucia Cerundolo ◽  
Mike Bunce ◽  
Dicken D.H. Koo ◽  
Kenneth I. Welsh ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Nested Pcr ◽  
Solid Organ Transplantation ◽  
Class Ii ◽  
Hla Class Ii ◽  
Solid Organ ◽  
Chain Reaction ◽  
Specific Primers ◽  
Polymerase Chain ◽  
Hla Class Ii Alleles

Abstract It is widely accepted that donor leukocytes survive within the recipient periphery after blood transfusion or solid organ transplantation. The significance of this microchimerism remains unclear, partially because of the insecurity of assays used to detect the donor-derived material. The techniques used to detect donor-derived DNA within recipient peripheral blood rely largely on major histocompatibility complex class II polymorphism. We and others have shown that the sensitivity of polymerase chain reaction with sequence-specific primers (PCR-SSP) typing for HLA class II alleles can be increased 100-fold by the addition of a primary amplification step (nested PCR-SSP). We have now extended this technique to encompass typing for HLA class I alleles, thereby adding flexibility to microchimerism testing by enabling testing of recipients HLA-DR matched with their donors. However, the high level of sensitivity achieved with the technique (1:100,000) leads to a concomitant decrease in the specificity that results in the amplification of unexpected products, a phenomenon we encountered in the development of our nested PCR-SSP typing system for HLA class II alleles. We describe here how it is possible to compensate for these anomalies by including multiple testing of a pretransfusion sample that acts as a specificity control, establishing a rigorous baseline for subsequent analysis.

Nested Polymerase Chain Reaction With Sequence-Specific Primers Typing for HLA-A, -B, and -C Alleles: Detection of Microchimerism in DR-Matched Individuals

Blood ◽  
1999 ◽  
Vol 94 (4) ◽  
pp. 1471-1477
Author(s):  
Anthony S. Carter ◽  
Lucia Cerundolo ◽  
Mike Bunce ◽  
Dicken D.H. Koo ◽  
Kenneth I. Welsh ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Nested Pcr ◽  
Solid Organ Transplantation ◽  
Class Ii ◽  
Hla Class Ii ◽  
Solid Organ ◽  
Chain Reaction ◽  
Specific Primers ◽  
Polymerase Chain ◽  
Hla Class Ii Alleles

It is widely accepted that donor leukocytes survive within the recipient periphery after blood transfusion or solid organ transplantation. The significance of this microchimerism remains unclear, partially because of the insecurity of assays used to detect the donor-derived material. The techniques used to detect donor-derived DNA within recipient peripheral blood rely largely on major histocompatibility complex class II polymorphism. We and others have shown that the sensitivity of polymerase chain reaction with sequence-specific primers (PCR-SSP) typing for HLA class II alleles can be increased 100-fold by the addition of a primary amplification step (nested PCR-SSP). We have now extended this technique to encompass typing for HLA class I alleles, thereby adding flexibility to microchimerism testing by enabling testing of recipients HLA-DR matched with their donors. However, the high level of sensitivity achieved with the technique (1:100,000) leads to a concomitant decrease in the specificity that results in the amplification of unexpected products, a phenomenon we encountered in the development of our nested PCR-SSP typing system for HLA class II alleles. We describe here how it is possible to compensate for these anomalies by including multiple testing of a pretransfusion sample that acts as a specificity control, establishing a rigorous baseline for subsequent analysis.

Evaluation of cytomegalovirus infection in low-birth weight children by breast milk using a real-time polymerase chain reaction assay

2015 ◽  
Vol 87 (5) ◽  
pp. 845-850 ◽  
Author(s):  
Romero-Gómez Maria Pilar ◽  
Cabrera Marta ◽  
Montes-Bueno María Teresa ◽  
Cendejas-Bueno Emilio ◽  
Segovia Cristina ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Birth Weight ◽  
Breast Milk ◽  
Low Birth Weight ◽  
Real Time ◽  
Polymerase Chain Reaction Assay ◽  
Cytomegalovirus Infection ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Low Birth Weight Children
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