Journal of Infection (2011) xx, 1e7

Enhancing the timeframe between blood collection and interferon gamma launch assay applying T-Cell XtendÒ J. J. M. Bouwman a, 2., S. Farrenheit. T. Thijsen a, A. W. Bossink b

a b

Division of Medical Microbiology and Immunology, Diakonessen Hospital Utrecht, The Netherlands Department of Pulmonology, Diakonessen Hospital, Utrecht, Holland

Accepted on the lookout for October 2011

T-Cell XtendÒ; TSPOT. TBÒ; Lithium heparin; IGRA; Interferon gamma; Tuberculosis


Summary Vacutainer CPT tubes need blood samples intended for TSPOT. TB to be refined within almost 8 h. In this study we evaluated the power of T-Cell Xtend to keep the number and performance of lymphocytes after twenty four and 48 h of blood storage, giving comparable test outcomes as in newly isolated specimens. Methods: Whole blood individuals from fifty nine individuals had been collected in Vacutainer CPT tubes (CPT) and lithium heparin (LH) tubes. CPT tubes were processed within just 8 they would. T-Cell Xtend was included in LH pontoons after twenty four or twenty four h. We all also left LH pontoons untreated to get 48 h. Total number of white blood cells (WBC) and amounts of lymphocytes and granulocytes were determined in the remote Peripheral Blood Mononuclear Cellular material (PBMC). We also evaluated the performance of T-Cell Xtend in the TSPOT. TB assay. Benefits: PBMC yields from T-Cell Xtend remedied LH examples did not differ from PBMC yields from CPT tubes, nevertheless T-Cell Xtend had a evident effect on the proportions of lymphocytes and granulocytes. The mean lymphocyte percentage in PBMCs separated from fresh CPT blood vessels was 84. 31 Æ 1 . 14% (at t Z 48 h), but was decreased to 52. seventy two Æ several. 34% (p < zero. 05) in untreated LH blood (at t Z 48 h). This result was neutralized by T-Cell Xtend (85. 44 Æ 0. 74%). We observed a similar although opposite impact on granulocytes: The mean proportion in untreated LH bloodstream was increased to 45. 9 Æ 3. 67% (p < 0. 001) compared to CPT blood (8. 26 Æ 0. 89%). Treatment of LH samples with T-Cell Xtend (48 h) restored the proportion of granulocytes to eight. 47 Æ 0. 61%. Enumeration of spots in the TSPOT. TB assay shown good arrangement between CPT and T-Cell Xtend benefits, even following 48 h.

Abbreviations: PBMC, Peripheral Blood Mononuclear Cellular material; CPT, Cell Preparation Conduit; LH, Li (symbol) Heparin; LTBI, Latent Tuberculosis Infection; IGRA, Interferon Molteplicit? Release Assay; MTB, Mycobacterium tuberculosis sophisticated; PPD, Purified Protein Offshoot; ESAT-6, Early on Secreted Antigenic Target-6; CFP10, Culture Filter Protein twelve; ELISA, Enzyme Linked Immuno Sorbent Assay; ELISPOT, Chemical Linked Immuno Spot; IFN-g, Interferon Gamma; PHA, Phytohaemagglutinin; PMN, Polymorphonuclear leukocytes; SFC, Spot forming cells; TST, Tuberculin Pores and skin Test; WBC, White Bloodstream Cells. 2. Corresponding writer. Department of Medical Microbiology and Immunology, Diakonessen Medical center Utrecht, Bosboomstraat 1, 3582 KE Utrecht, The Netherlands. Tel.: þ31 88 250 6075; fax: þ31 88 two hundred fifty 6695. E-mail address: [email protected] nl (J. J. M. Bouwman). 0163-4453/$36 ª 2011 The United kingdom Infection Affiliation. Published simply by Elsevier Limited. All legal rights reserved. doi: 10. 1016/j. jinf. 2011. 10. 017 Please refer to this article in press as: Bouwman JJM, et al., Improving the timeframe between bloodstream collection and interferon molteplicit? release assay using T-Cell XtendÒ, M Infect (2011), doi: 10. 1016/j. jinf. 2011. 10. 017


J. T. M. Bouwman et 's.

Conclusions: T-Cell Xtend efficiently removes granulocytes from PBMC suspensions and increases the portion of lymphocytes. TSPOT. TB results from T-Cell Xtend treated blood samples have reached least comparable to the benefits obtained from the current CPT approach. Use of common lithium heparin blood coupled with T-Cell Xtend allows up to 48 they would storage of blood samples to get batched finalizing and may additional decrease the charge of indeterminate TSPOT. TB results. ª 2011 The British Infection Association. Published by Elsevier Ltd. All rights arranged.


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