Diagnostic laboratory standardization and validation of platelet transmission electron microscopy

Dong Chen, Cindy B. Uhl, Sandra C. Bryant, Marcy Krumwiede, Ryan L. Barness, Mary C. Olson, Susan C. Gossman, Sibel Erdogan Damgard, Scott I. Gamb, Lisa A. Cummins, Jon E. Charlesworth, Christina M. Wood-Wentz, Jeffrey L Salisbury, Elizabeth A. Plumhoff, Elizabeth M. Van Cott, Rong He, Deepti M. Warad, Rajiv K. Pruthi, John A. Heit, William L. Nichols & 1 others James G. White

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Platelet transmission electron microscopy (PTEM) is considered the gold standard test for assessing distinct ultrastructural abnormalities in inherited platelet disorders (IPDs). Nevertheless, PTEM remains mainly a research tool due to the lack of standardized procedures, a validated dense granule (DG) count reference range, and standardized image interpretation criteria. The aim of this study was to standardize and validate PTEM as a clinical laboratory test. Based on previously established methods, we optimized and standardized preanalytical, analytical, and postanalytical procedures for both whole mount (WM) and thin section (TS) PTEM. Mean number of DG/platelet (plt), percentage of plts without DG, platelet count (PC), mean platelet volume (MPV), immature platelet fraction (IPF), and plt light transmission aggregometry analyses were measured on blood samples from 113 healthy donors. Quantile regression was used to estimate the reference range for DG/plt, and linear regression was used to assess the association of DG/plt with other plt measurements. All PTEM procedures were standardized using commercially available materials and reagents. DG interpretation criteria were established based on previous publications and expert consensus, and resulted in improved operator agreement. Mean DG/plt was stable for 2 days after blood sample collection. The median within patient coefficient of variation for mean DG/plt was 22.2%; the mean DG/plt reference range (mid-95th %) was 1.2–4.0. Mean DG/plt was associated with IPF (p = .01, R2 = 0.06) but not age, sex, PC, MPV, or plt maximum aggregation or primary slope of aggregation (p > .17, R2 < 0.02). Baseline ultrastructural features were established for TS-PTEM. PTEM was validated using samples from patients with previously established diagnoses of IPDs. Standardization and validation of PTEM procedures and interpretation, and establishment of the normal mean DG/plt reference range and PTEM baseline ultrastructural features, will facilitate implementation of PTEM as a valid clinical laboratory test for evaluating ultrastructural abnormalities in IPDs.

Original languageEnglish (US)
Pages (from-to)1-9
Number of pages9
JournalPlatelets
DOIs
StateAccepted/In press - Jun 2 2018

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Transmission Electron Microscopy
Blood Platelets
Reference Values
Mean Platelet Volume
Platelet Count

Keywords

  • Electron microscopy
  • inherited platelet disorders
  • platelet (plt)

ASJC Scopus subject areas

  • Hematology

Cite this

Chen, D., Uhl, C. B., Bryant, S. C., Krumwiede, M., Barness, R. L., Olson, M. C., ... White, J. G. (Accepted/In press). Diagnostic laboratory standardization and validation of platelet transmission electron microscopy. Platelets, 1-9. https://doi.org/10.1080/09537104.2018.1476682

Diagnostic laboratory standardization and validation of platelet transmission electron microscopy. / Chen, Dong; Uhl, Cindy B.; Bryant, Sandra C.; Krumwiede, Marcy; Barness, Ryan L.; Olson, Mary C.; Gossman, Susan C.; Erdogan Damgard, Sibel; Gamb, Scott I.; Cummins, Lisa A.; Charlesworth, Jon E.; Wood-Wentz, Christina M.; Salisbury, Jeffrey L; Plumhoff, Elizabeth A.; Van Cott, Elizabeth M.; He, Rong; Warad, Deepti M.; Pruthi, Rajiv K.; Heit, John A.; Nichols, William L.; White, James G.

In: Platelets, 02.06.2018, p. 1-9.

Research output: Contribution to journalArticle

Chen, D, Uhl, CB, Bryant, SC, Krumwiede, M, Barness, RL, Olson, MC, Gossman, SC, Erdogan Damgard, S, Gamb, SI, Cummins, LA, Charlesworth, JE, Wood-Wentz, CM, Salisbury, JL, Plumhoff, EA, Van Cott, EM, He, R, Warad, DM, Pruthi, RK, Heit, JA, Nichols, WL & White, JG 2018, 'Diagnostic laboratory standardization and validation of platelet transmission electron microscopy', Platelets, pp. 1-9. https://doi.org/10.1080/09537104.2018.1476682
Chen, Dong ; Uhl, Cindy B. ; Bryant, Sandra C. ; Krumwiede, Marcy ; Barness, Ryan L. ; Olson, Mary C. ; Gossman, Susan C. ; Erdogan Damgard, Sibel ; Gamb, Scott I. ; Cummins, Lisa A. ; Charlesworth, Jon E. ; Wood-Wentz, Christina M. ; Salisbury, Jeffrey L ; Plumhoff, Elizabeth A. ; Van Cott, Elizabeth M. ; He, Rong ; Warad, Deepti M. ; Pruthi, Rajiv K. ; Heit, John A. ; Nichols, William L. ; White, James G. / Diagnostic laboratory standardization and validation of platelet transmission electron microscopy. In: Platelets. 2018 ; pp. 1-9.
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AU - Pruthi, Rajiv K.

AU - Heit, John A.

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