適用樣本類(lèi)型廣泛 

可常溫保存樣本 

穩(wěn)定長(zhǎng)達(dá)14天 

高質(zhì)量流式分析必備

CYTOMARK公司簡(jiǎn)介

Caltag Medsystems 是 Cytomark 的母公司，由首席執(zhí)行官 Tim Almond 博士于 2001 年創(chuàng)建。TransFix®是由英國(guó)國(guó)家衛(wèi)生服務(wù)(NHS)下屬機(jī)構(gòu) NEQAS 的醫(yī)學(xué)科學(xué)家于 20 世紀(jì) 90 年代開(kāi)發(fā)的。2005 年，Tim Almond 博士推出了 Cytomark，并獲得了 NHS 許可的 TransFix®，使其易于為世界各地的臨床和科學(xué)機(jī)構(gòu)使用。

什么是TransFix?

 

TransFix®是一種穩(wěn)定試劑，保存細(xì)胞抗原，防止細(xì)胞降解的各種標(biāo)本類(lèi)型的流式細(xì)胞分析。

也可用于各種標(biāo)本類(lèi)型，包括:腦脊液，淋巴結(jié)，骨髓，動(dòng)物(小鼠、綿羊、雞、海洋生物)血液，循環(huán)腫瘤細(xì)胞，細(xì)胞游離RNA和外泌體分離。

TransFix用途廣泛——多功能穩(wěn)定劑

細(xì)胞與細(xì)胞表面抗原——穩(wěn)定細(xì)胞形態(tài)與表面抗原長(zhǎng)達(dá)14D

核酸（細(xì)胞內(nèi)核酸與ctDNA/RNA）——可穩(wěn)定樣品中循環(huán)無(wú)細(xì)胞核酸

外泌體與外泌體標(biāo)志物——滿(mǎn)足對(duì)外泌體TEM投射檢驗(yàn)的需求，并且穩(wěn)定多種外泌體microRNA(10D后也可滿(mǎn)足提取分析標(biāo)志物)

CTC細(xì)胞——已在對(duì)腫瘤、心血管疾病與其他疾病的標(biāo)志物的研究成功多天穩(wěn)定樣本，滿(mǎn)足大量樣本同時(shí)檢測(cè)

細(xì)胞樣本穩(wěn)定難點(diǎn)

1.     生物樣本中細(xì)胞隨時(shí)間降解，影響檢測(cè)質(zhì)量并增加檢測(cè)人員的工作壓力

2.     高通量大樣本難以同時(shí)檢測(cè),不同地點(diǎn)儀器不同時(shí)間與檢測(cè)人員間的進(jìn)行測(cè)試的誤差難以消除

3.     部分產(chǎn)品只針對(duì)細(xì)胞抗原或核酸進(jìn)行穩(wěn)定，難以滿(mǎn)足多種下游檢測(cè)的需求，增加多次采樣的壓力

4.     珍惜樣本采樣難度大，檢測(cè)技術(shù)要求高產(chǎn)生的樣本保存時(shí)間與無(wú)法及時(shí)檢測(cè)的矛盾

外泌體研究中血小板隨時(shí)間釋放外泌體與短片段靶標(biāo)易降解的難點(diǎn)

為什么要選擇TransFix——獨(dú)特優(yōu)勢(shì)

TransFix比其他細(xì)胞固定與穩(wěn)定產(chǎn)品相比較有著無(wú)法替代的多種優(yōu)勢(shì)

1.TransFix可適用于多種樣品，并根據(jù)您不同的下游試驗(yàn)需求調(diào)整用量（詳情請(qǐng)參照廠家的使用指南）

可適用于：

*  全血樣品（包括表面抗原、紅細(xì)胞、血小板、CTC細(xì)胞與ctDNA/RNA等）

* CSF腦脊液

*  骨髓樣本及其他細(xì)針吸出樣本

*  肺泡灌洗液

*  間充質(zhì)干細(xì)胞與胚胎細(xì)胞

*  其他文獻(xiàn)中應(yīng)用的動(dòng)物及組織等樣本

2.     TransFix在2-8℃下可保存全血樣本14天，常溫保存細(xì)胞長(zhǎng)達(dá)3-5天

3.     TransFix不僅可以保存細(xì)胞與表面抗原，在與不同科學(xué)家合作中開(kāi)發(fā)了其更多的用途

*  減少細(xì)胞與細(xì)胞表面抗原的降解

*  穩(wěn)定全血樣本中血小板與外泌體包含多種外泌體microRNA生物靶標(biāo)

*  穩(wěn)定CTC細(xì)胞與ctDNA/RNA

*  穩(wěn)定細(xì)胞或樣本中核酸（qpcr前需要提取核酸后再進(jìn)行）

TransFix應(yīng)用場(chǎng)景

全血樣本的 14 天流式細(xì)胞對(duì)比分析

白細(xì)胞亞群是根據(jù)它們的光散射譜和細(xì)胞表面抗原通過(guò)流式細(xì)胞儀區(qū)分的。這些亞群的數(shù)量變化使血液系統(tǒng)惡性腫瘤(如白血病)的鑒別診斷和監(jiān)測(cè)成為可能，并使艾滋病毒/艾滋病患者的遠(yuǎn)程免疫監(jiān)測(cè)成為可能。

傳統(tǒng)采血管采集樣本后必須在靜脈穿刺后 48 小時(shí)內(nèi)對(duì)血液樣本進(jìn)行流式細(xì)胞分析。且在老年血液樣本表現(xiàn)出難以區(qū)分的細(xì)胞亞群和不準(zhǔn)確的絕對(duì)細(xì)胞計(jì)數(shù)，這可能導(dǎo)致錯(cuò)誤的臨床結(jié)果。

而采用含有 Trans Fix 的真空采血管進(jìn)行樣本采集與貯存后，可以看到 14天后的流式細(xì)胞分析結(jié)果仍呈現(xiàn)準(zhǔn)確的細(xì)胞亞群統(tǒng)計(jì)結(jié)果。

與新鮮血液相比 TVTs 在第 15 天顯示等效的白細(xì)胞譜

TVT 穩(wěn)定的血液樣本在第 15 天顯示出與采用流式細(xì)胞術(shù)金標(biāo)準(zhǔn)采樣管（BD K3EDTA 真空采血管）新鮮血液樣本有相似的白細(xì)胞譜，細(xì)胞碎片水平低，CD3, CD4, CD8, CD16+56, CD45 和 CD19 群體分離良好，平均熒光強(qiáng)度與新鮮血液相似，使白細(xì)胞亞群分離清楚。以下數(shù)據(jù)展示使用該等標(biāo)記之 HIV患者的流式細(xì)胞術(shù)點(diǎn)圖。

相關(guān)文獻(xiàn)

1.         Lysák et al (2010) Interlaboratory variability of CD34+ stem cell enumeration. A pilot study to national external quality assessment within the Czech Republic. Int. Jnl. Lab. Hem. 32, e229–e236. https://doi.org/10.1111/j.1751-553X.2010.01244.x

2.         Levering et al. (2007) Flow Cytometric CD34+ Stem Cell Enumeration: Lessons from Nine Years’ External Quality Assessment Within the Benelux Countries. Cytometry Part B (Clinical Cytometry) 72B: 178–188. https://doi.org/10.1002/cyto.b.20351

3.         Thastrup et al. (2019) Flow cytometric detection of leukemic blasts in cerebrospinal fluid predicts risk of relapse in childhood acute lymphoblastic leukemia: a Nordic Society of Pediatric Hematology and Oncology study. Leukemia 34: 336–346. https://doi.org/10.1038/s41375-019-0570-1

4.         Levinsen et al (2016) Leukemic blasts are present at low levels in spinal fluid in one-third of childhood acute lymphoblastic leukemia cases. Pediatr Blood Cancer 63; 1935–1942. https://doi.org/10.1002/pbc.26128

5.         Beiral et al. (2014) The impact of stem cells on electron fluxes, proton translation, and ATP synthesis in kidney mitochondria after ischemia/reperfusion. Cell Transplant 23(2): 207-20. https://doi.org/10.3727/096368912X659862

6.         Harrison D, Ward R, Bastow S, Parr A, Macro S, and Wallace PK. Interlaboratory Comparison of the TransFix®/EDTA Vacuum Blood Collection Tube with the 5 mL Cyto‐Chex® BCT. Cytometry Part B 2018; 9999: 1–12.

7.         Leligdowicz et al. Direct Relationship between Virus Load and Systemic Immune Activation in HIV-2 Infection. J Infect Dis 201 (1): 114-122. https://doi.org/10.1086/648733

8.         Olivares et al. (2014) Double Blind, Randomised, Placebo-Controlled Intervention Trial to Evaluate the Effects of Bifido Longum CECT 7347 in Children with Newly Diagnosed Coeliac Disease. British Journal of Nutrition 112 (1); 30-40. https://doi.org/10.1017/S0007114514000609

9.         Poirier et al. (2016) First-in-Human Study in Healthy Subjects with FR104, a Pegylated Monoclonal Antibody Fragment Antagonist of CD28. J Immunol 197(12): 4593-4602.  https://doi.org/10.4049/jimmunol.1601538

10.     de Jongste AH, Kraan J, van den Broek PD, Brooimans RA, Bromberg JE, van Montfort KA, Smitt PAS, and Gratama JW. Use of TransFix™ Cerebrospinal Fluid Storage Tubes Prevents Cellular Loss and Enhances Flow Cytometric Detection of Malignant Hematological Cells After 18 Hours of Storage. Cytometry Part B 2014; 86B: 272– 279. https://doi.org/10.1002/cyto.b.21097

11.     Kaenzig et al. Evaluation of TransFix/EDTA CSF Sample Storage Tubes compared to alternative preservation methods (Caltag Medsystems Ltd 2022- unpublished)

12.     Johansson U, Bloxham D, Couzens S, Jesson J, Morilla R, Erber W, and Macey M. (2014). Guidelines on the use of multicolour flow cytometry in the diagnosis of haematological neoplasms. British Journal of Haematology, 165, 455–488.

13.     Baraniskin A, Deckert M, Schulte-Altedorneburg G, Schlegel U, Schroers R. Current strategies in the diagnosis of diffuse large B-cell lymphoma of the central nervous system. Br J Haematol. 2012 Feb;156(4):421-32. doi: 10.1111/j.1365-2141.2011.08928

14.     Del Principe MI, Gatti A, Johansson U, Buccisano F, Brando B. ESCCA/ISCCA protocol for the analysis of cerebrospinal fluid by multiparametric flow‐cytometry in hematological malignancies. Cytometry. 2020; 1– 13. https://doi.org/10.1002/cyto.b.21981

15.     Peñalver FJ, Sancho JM, de la Fuente A, Olave MT, Martín A, Panizo C, Pérez E, Salar A, Orfao A. Guidelines for diagnosis, prevention and management of central nervous system involvement in diffuse large B-cell lymphoma patients by the Spanish Lymphoma Group (GELTAMO). Haematologica 2017;102(2):235-245; https://doi.org/10.3324/haematol.2016.149120

16.     Quijano S, López A, Manuel Sancho J, Panizo C, Debén G, Castilla C, Antonio García-Vela J, Salar A, Alonso-Vence N, González-Barca E, Peñalver FJ, Plaza-Villa J, Morado M, García-Marco J, Arias J, Briones J, Ferrer S, Capote J, Nicolás C, Orfao A; Spanish Group for the Study of CNS Disease in NHL. Identification of leptomeningeal disease in aggressive B-cell non-Hodgkin’s lymphoma: improved sensitivity of flow cytometry. J Clin Oncol. 2009 Mar 20;27(9):1462-9. doi: 10.1200/JCO.2008.17.7089.

17.     Correia RP, Bortolucci ACA, Lopes ACW, Sandes AF, Azambuja AP, Viana MA, et al. Recommendations for quality assurance in multiparametric flow cytometry: first concensus of the Brazilian Group of Flow Cytometry (GBCFLUX). J. Bras. Patol. Med. Lab.2015;51(6):389-396 https://doi.org/10.5935/1676-2444.20150061

18.     Kraan J, Gratama JW, Haioun C, Orfao A, Plonquet A, Porwit A, Quijano S, Stetler-Stevenson M, Subira D and Wilson W. (2008), Flow Cytometric Immunophenotyping of Cerebrospinal Fluid. Current Protocols in Cytometry, 45: 6.25.1-6.25.16. https://doi.org/10.1002/0471142956.cy0625s45

Campos A, Trujillo L, López D, Beltrán L, Arias E, Vélez G, Infante A, Reyes I, Vizcaíno M, Guzmán P C, Herrera MV, Solano J, Londono D, Cañas A, Pretelt F, Pérez JC, Cardozo C, Fiorentino S, and Quijano, S. (2017). Study of body fluid samples using flow cytometry: Six years of experience at the Hospital Universitario San Ignacio – Pontificia Universidad Javeriana, Bogota – Colombia. Universitas Scientiarum (2017). 22. 123-143. 10.11144/Javeriana.SC22-2.sobf.