Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms.

Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. Research Abstract Details 

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Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. Abstract Text:

Tzong-Hae Lee,Daniel M Chafets,William Reed,Li Wen,Yunting Yang,Jennifer Chen,Garth H Utter,John T Owings,Michael P Busch,

BACKGROUND: The characterization of microchimerism (MC) by gene amplification has been limited by few allogeneic markers, ascertainment bias, and assay analytic performance. To address this, a panel of 12 MC assays based on insertion-deletion (InDel) polymorphisms had been optimized. STUDY DESIGN AND METHODS: The InDel assays were validated with comprehensive in vitro spiking studies at the stochastic limit of detection. Their ability was also determined to ascertain MC of unknown source genotype with both theoretical and actual donor-recipient pairs, and the assays were applied to a clinical population of 73 trauma patients who received transfusions where MC was previously characterized by HLA-based assays alone. RESULTS: In the stochastic spiking experiments, all assays were sensitive to a single copy of target DNA, and no false-positive amplification occurred among 1128 samples studied. Among 219 theoretical donor-recipient pairs, informative alleles existed for 99.5 percent with both InDel and HLA compared to 91.3 percent with HLA alone. In the clinical population, 33 cases of MC were detected (9 more cases than by HLA-DR alone) in the nonleukoreduced (non-LR) group and 8 cases (1 more case than by HLA-DR) in the LR group for the short-term follow-up. Among 27 long-term follow-up samples, 8 cases were detected overall (3 more cases than by HLA-DR alone). CONCLUSION: It is concluded that an InDel-based assay panel has excellent technical performance characteristics while also allowing for ascertainment of some MC cases not detectable with HLA alone. The tandem use of both the InDel and the HLA provides a powerful tool for the enhanced ascertainment of MC.

Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. Publishing Authors By Initials

TH Lee,DM Chafets,W Reed,L Wen,Y Yang,J Chen,GH Utter,JT Owings,MP Busch,

For similar disorders of environmental origin: wounds and injuries research abstracts see: disorders of environmental origin: wounds and injuries research

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Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Transfusion

VOLUME: 46

Page Numbers: 1870-8

Journal Abbreviation: Transfusion

ISSN: 0041-1132

DAY: 3

MONTH: Nov

YEAR: 2006

Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 417360

Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. Keywords Mesh Terms:

KEYWORDS: Wounds and Injuries

MESH TERMS: therapy

Chemical & Substance for Abstract: Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. Information

Substance Name: HLA-DR Antigens

Registry Number: 0

Grant and Affiliation Information for Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms.

AFFILIATION: Blood Systems Research Institute, San Francisco, California 94118, USA. tlee@bloodsystems.org

Country: United States

AGENCY: United States NHLBI

GRANT: P50 HL54476

ACRONYM: HL

MEDLINETA: Transfusion

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