Real time-PCR assay estimating the naive T-cell pool in whole blood and dried blood spot samples: pilot study in young adults.

Author(s): Lang PO, Mitchell WA, Govind S, Aspinall R

Publication: J Immunol Methods, 2011, Vol. 369, Page 133-40

PubMed ID: 21619883 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of using DNA from PBMC versus DBS for the calculation of sj-TREC and T-cells counts via real time qPCR and flow cytometric analysis. Isolated PBMC were stored at -80 C until DNA extraction while DBS were stored desiccated at -20 degrees C.

   Summary of Findings:

   Leukocyte counts and sj-TREC/T-cells ratios were comparable when calculated by qPCR analysis or measured by flow cytometry. Further, both DNA extracted from PBMC and from DBS fell into an acceptable purity range, and qPCR results using DNA from each specimen type were not significantly different.

   Biospecimens

   • Bodily Fluid - Blood
   • Cell - Blood

   Preservative Types

   • Frozen
   • Other Preservative

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Flow cytometry
   DNA	Real-time qPCR
   DNA	Spectrophotometry
   Cell count/volume	Real-time qPCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Air-dried Frozen
   Real-time qPCR Specific	Targeted nucleic acid	Albumin sj-TREC VD junction of T-cell receptor-beta
   Flow cytometry Specific	Technology platform	Real time qPCR

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