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Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms.
Transfusion. 2006 Nov; 46(11):1870-8.T

Abstract

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.

Authors+Show Affiliations

Blood Systems Research Institute, San Francisco, California 94118, USA. tlee@bloodsystems.orgNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Randomized Controlled Trial
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17076840

Citation

Lee, Tzong-Hae, et al. "Enhanced Ascertainment of Microchimerism With Real-time Quantitative Polymerase Chain Reaction Amplification of Insertion-deletion Polymorphisms." Transfusion, vol. 46, no. 11, 2006, pp. 1870-8.
Lee TH, Chafets DM, Reed W, et al. Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. Transfusion. 2006;46(11):1870-8.
Lee, T. H., Chafets, D. M., Reed, W., Wen, L., Yang, Y., Chen, J., Utter, G. H., Owings, J. T., & Busch, M. P. (2006). Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. Transfusion, 46(11), 1870-8.
Lee TH, et al. Enhanced Ascertainment of Microchimerism With Real-time Quantitative Polymerase Chain Reaction Amplification of Insertion-deletion Polymorphisms. Transfusion. 2006;46(11):1870-8. PubMed PMID: 17076840.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enhanced ascertainment of microchimerism with real-time quantitative polymerase chain reaction amplification of insertion-deletion polymorphisms. AU - Lee,Tzong-Hae, AU - Chafets,Daniel M, AU - Reed,William, AU - Wen,Li, AU - Yang,Yunting, AU - Chen,Jennifer, AU - Utter,Garth H, AU - Owings,John T, AU - Busch,Michael P, PY - 2006/11/2/pubmed PY - 2006/12/29/medline PY - 2006/11/2/entrez SP - 1870 EP - 8 JF - Transfusion JO - Transfusion VL - 46 IS - 11 N2 - 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. SN - 0041-1132 UR - https://www.unboundmedicine.com/medline/citation/17076840/Enhanced_ascertainment_of_microchimerism_with_real_time_quantitative_polymerase_chain_reaction_amplification_of_insertion_deletion_polymorphisms_ L2 - https://doi.org/10.1111/j.1537-2995.2006.00992.x DB - PRIME DP - Unbound Medicine ER -