Quantitative polymerase chain reaction technology in chimerism monitoring after hematopoietic stem cell transplantation: One center experience

HLA ◽  
2019 ◽  
Vol 94 (S2) ◽  
pp. 16-20
Author(s):  
Katarina Stingl Jankovic ◽  
Marija Maskalan ◽  
Marija Burek Kamenaric ◽  
Renata Zunec ◽  
Nadira Durakovic ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Stem Cell ◽  
Hematopoietic Stem Cell Transplantation ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Hematopoietic Stem ◽  
Polymerase Chain Reaction Technology ◽  
Polymerase Chain

scholarly journals Comparison of serology, antigenemia assay and the polymerase chain reaction for monitoring active cytomegalovirus infections in hematopoietic stem cell transplantation patients

2006 ◽  
Vol 48 (5) ◽  
pp. 275-278 ◽  
Author(s):  
Sandra Helena Alves Bonon ◽  
Cláudio Lúcio Rossi ◽  
Cármino Antonio de Souza ◽  
Afonso Celso Vigorito ◽  
Sandra Cecília Botelho Costa
Keyword(s):  
Polymerase Chain Reaction ◽  
Stem Cell ◽  
Hematopoietic Stem Cell Transplantation ◽  
Stem Cell Transplantation ◽  
Median Time ◽  
Igg Antibodies ◽  
Chain Reaction ◽  
Hematopoietic Stem ◽  
Antigenemia Assay ◽  
Polymerase Chain

Forty-six allogeneic hematopoietic stem cell transplantation (HSCT) patients were monitored for the presence of CMV antibodies, CMV-DNA and CMV antigens after transplantation. Immunoenzymatic serological tests were used to detect IgM and the increase in CMV IgG antibodies (increase IgG), a nested polymerase chain reaction (N-PCR) was used to detect CMV-DNA, and an antigenemia assay (AGM) was used to detect CMV antigens. The presence of CMV-IgM and/or CMV-increase IgG antibodies was detected in 12/46 (26.1%) patients, with a median time between HSCT and the detection of positive serology of 81.5 days. A positive AGM was detected in 24/46 (52.2%) patients, with a median time between HSCT and antigen detection of 62 days. Two or more consecutive positive N-PCR results were detected in 32/46 (69.5%) patients, with a median time between HSCT and the first positive PCR of 50.5 days. These results confirmed that AGM and mainly PCR are superior to serology for the early diagnosis of CMV infection. Six patients had CMV-IgM and/or CMV-increase IgG with a negative AGM (five cases) or N-PCR assay (one case). In five of these cases the serological markers were detected during the first 100 days after HSCT, the period of highest risk. These findings support the idea that serology may be useful for monitoring CMV infections in HSCT patients, especially when PCR is unavailable.

Quantitative assessment of hematopoietic chimerism after bone marrow transplantation by real-time quantitative polymerase chain reaction

Blood ◽  
2002 ◽  
Vol 99 (12) ◽  
pp. 4618-4625 ◽  
Author(s):  
Mehdi Alizadeh ◽  
Marc Bernard ◽  
Bruno Danic ◽  
Charly Dauriac ◽  
Brigitte Birebent ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Stem Cell ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Assessment ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Hematopoietic Stem ◽  
Pcr Method ◽  
Polymerase Chain

We have developed a real-time quantitative polymerase chain reaction (PCR) assay using TaqMan technology (Applied Biosystems, Foster City, CA) for monitoring donor cell engraftment in allogenic hematopoietic stem cell transplant recipients. For this purpose, we selected 19 specific sequence polymorphisms belonging to 11 human biallelic loci located on 9 different chromosomes. Using a set of specially designed primers and fluorogenic probes, we evaluated the 19 markers' informativity on a panel of 126 DNA samples from 63 recipient/donor pairs. In more than 90% of these pairs, discrimination between recipient and donor genetic profile was possible. By using serial dilutions of mixed DNAs, we evaluated the linearity and sensitivity of the method. A linear correlation with rhigher than 0.98 and a sensitivity of 0.1% proved reproducible. Fluorescent-based PCR of short tandem repeats (STR-PCR) and real-time PCR chimerism assay were compared with a panel of artificial cell mixtures. The main advantage of the real-time PCR method over STR-PCR chimerism assays is the absence of PCR competition and plateau biases, and results evidenced greater sensitivity and linearity with the real-time PCR method. Furthermore, different samples can be tested in the same PCR run with a final result in fewer than 48 hours. Finally, we prospectively analyzed patients who received allografts and present 4 different clinical situations that illustrate the informativity level of our method. In conclusion, this new assay provides an accurate quantitative assessment of mixed chimerism that can be useful in guiding early implementation of additional treatments in hematopoietic stem cell transplantation.

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