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Effects of EDTA on Routine and Specialized Coagulation Testing

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Effects of EDTA on Routine and Specialized Coagulation Testing 27Laboratory HematologyAbroun 15:45-4818:27-38 et al © 20102012 Carden Jennings Publishing Co., Ltd. doi: 10.1532/LH96.0901310.1532/LH96.11003 BiologyEffects of and EDTA Bioinformatics on Routine of and Myeloma Specialized Cell SaeidCoagulation Abroun,1 Najmaldin Testing Saki, and2 Rahim an Fakher, Easy3 FarahnazMethod Asghari to Distinguish4 EDTA-Treated1Department of Hematology and Bloodfrom Banking, Citrated School of Medical Sciences,Plasma Tarbiat ModaresSamples University, Tehran, Iran; 2Research Center of Thalassemia & Hemoglobinopathy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 3Physiology Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; 4Faculty of Medicine, University Rondaof Rostock, A.Rostock, Crist, Germany1,2 Kathie Gibbs,2 George M. Rodgers,1-4 Kristi J. Smock1,2,4 Received May 14, 2011; December 13, 2011, February 10, 2012; August 7, 2012 1ARUP Institute for Clinical and Experimental Pathology, Salt Lake City; 2ARUP Hemostasis/Thrombosis Laboratory, ⌦Salt Lake City; Departments of 3Medicine and 4Pathology, University of Utah, Salt Lake City, Utah, USA Received October 7, 2009; received in revised form November 5, 2009; accepted November 6, 2009 ABSTRACT approximately 14,600 new cases of myeloma are diagnosed ABSTRACT each year in the United States and that it develops in 1 to Multiple myeloma (MM) is a plasma cell disorder that 4 people per 100,000 people each year. With conventional occurs in about 10% of all hematologic cancers. The major- treatment, the prognosis is 3 to 4 years, which may be Coagulation testing is performed with citrate-treated ity of patients (99%) are over 50 years of age when diag- treatedextended with to 5citrate, to 7 yearswhich with anticoagulates advanced treatments. by chelating MM ◆cal is- plasma. Samples submitted in other anticoagulants, such nosed. In the bone marrow (BM), stromal and hematopoi- ciumthe second ions necessarymost prevalent for calcium-dependent blood cancer (10%) coagulation after non- as EDTA, should not be tested. We aimed to evaluate the etic stem cells (HSCs) are responsible for the production of reactionsHodgkin’s [1,2]. lymphoma Calcium (NHL). is replenished It represents by the approximately test reagents effects of EDTA on routine and specialized coagulation blood cells. Therefore any destruction or/and changes within at1% the of timeall cancers of testing. and 2%EDTA of all is canceran anticoagulant deaths. Although that che its- tests and to establish sodium tetraphenylborate testing as a the BM undesirably impacts a wide range of hematopoiesis, latespeak calciumage of onset irreversibly, is from 65a quality to 70 yearsthat hasof age, the recentpotential statis to- quick and reliable method to identify EDTA-treated plasma causing diseases and influencing patient survival. In order dramaticallytics indicate bothaffect an the increasing results and incidence interpretation and an earlierof coagu age- samples. We performed the following measurements on to establish an effective therapeutic strategy, recognition lationof onset tests [1]. [3-8].Usually, EDTA MM issamples diagnosed must by bloodbe identified sample tests and citrate- and EDTA-treated plasma samples from 10 healthy of the biology and evaluation of bioinformatics models for rejectedincluding by peripheral the coagulation blood smear laboratory and protein to avoid electrophore negative- volunteers: sodium tetraphenylborate testing, prothrombin myeloma cells are necessary to assist in determining suitable impactssis, but inon somepatient cases care. microscopic In the present examination study, we of aimed the BM to time (PT), partial thromboplastin time (PTT), potassium methods to cure or prevent disease complications in patients. evaluatebiopsy and the x-rays effects of bonesof EDTA were alsoon routinegood indicators and specialized of MM. concentration, and functional assays for factors II, V, VII, This review presents the evaluation of molecular and cellular coagulationIn 2003, the tests International and to describe Myeloma a new Working method Group for identify agreed- VIII, IX, X, XI, XII, proteins C and S, and antithrom- aspects of MM such as genetic translocation, genetic analysis, ingon aEDTA-treated diagnostic criteria plasma for samples. symptomatic and asymptomatic bin. Mean values for citrate- and EDTA-treated plasma cell surface marker, transcription factors, and chemokine myeloma,In our referencewhich was laboratory subsequently setting, updated we do in not2009: receive The were most different for PT, PTT, factors V and VIII, and signaling pathways. It also briefly reviews some of the mecha- ◆presenceprimary-draw of unexplained tubes, making anemia it difficultby hemoglobin to discern < 10 whether g/dL, proteins C and S. Sodium tetraphenylborate testing cor- nisms involved in MM in order to develop a better under- thekidney correct dysfunction anticoagulant follow wasby raisedused. serumAs part creatinine, of our quality- a high rectly classified 100% of citrate-treated and EDTA-treated standing for use in future studies. ◆erythrocyteassurance plan, sedimentation we measure rate, the and prothrombin a high serum time protein (PT) samples. We confirm that EDTA has effects on coagula- and> 30g/L, the partial which thromboplastin especially raised time immunoglobulin (PTT) on all samples. (Ig) in tion assays. Sodium tetraphenylborate testing is a quick, KEY WORDS: Multiplemyeloma◆◆•◆◆Chemokines◆◆•◆◆ Thisserum process and/or highallows proteinurea, us to detect which erroneously indicate Bence submitted Jones simple, and inexpensive method for coagulation laborato- serumprotein samples,in urine, whichhyper calcemiahave markedly > 2.75 prolongedmmol/L, infection, PT and ries to identify samplesSignaling erroneouslypathways◆◆•◆◆Bioinformatics submitted in EDTA. PTTbone valuespain by (often lytic lesions>150 seconds) or osteoporosis but correct with to compression within the Lab-Hematol. 2009;15:45-48. normalfractures, range neurological in a 1:1 mixingsymptom study. by hypercalcemia,This approach ordoes spinal not INTRODUCTION reliablycord damage identify by backEDTA-containing bone fracture. plasmaMM can samples, also be which diag- KEY WORDS: Citrate◆◆•◆◆EDTA◆◆•◆◆Coagulation usuallynosed if demonstratethe population prolonged of plasma (but cells measurable)in the BM biopsy PT and are Multiple myelomatesting◆◆•◆◆Preanalytical (MM) is a B-lineage cell variables malignancy PTT> 10% values [2,3]. that Based often on Internationaldo not correct Staging in a 1:1 System mixture (ISS) [9]. or characterized by the monoclonal expansion of malignant SuspectedDurie-Salmon EDTA-containing staging guidance, plasma there canare 3be classified confirmed stages by plasma cells within the bone marrow (BM). There are undetectabilityof MM (Table) of[4,5]. calcium (due to avid calcium chelation by INTRODUCTIONapproximately 45,000 people in the United States diagnosed EDTA)It is andlikely increased that MM potassium has evolved values from(EDTA pre-malignant tubes usually with MM, and the American Cancer Society estimates that containstages of EDTA clonal asplasma a potassium cell proliferation salt) [1,6,8-10]. termed Conducting monoclo- Coagulation testing is performed on plasma samples thesenal gammopathy chemical tests of is undetermineddisruptive to our significance laboratory (MGUS).work flow Correspondence and reprint requests: Saeid Abroun, Department of becausePatients theymay arebe notdiagnosed available with at theMGUS coagulation if they bench. fulfill We the HematologyCorrespondence and and Blood reprint Banking, requests: School Kristi of J.Medical Smock, Sciences,ARUP Labo Tar- havefollowing found 3 thatcriteria: testing 1) patientA monoclonal samples withparaprotein sodium tetraband- biatratories, Modares 500 ChipetaUniversity, Way, Tehran, Salt LakeIran; City,+98 21UT 82883860; 84108, USA fax: (e-mail:+98 21 phenylborateless than 30 g/Lis a simple(< 3 g/dL); and reliable 2) Plasma way cellsto identify less than EDTA- 10% [email protected]). (e-mail: [email protected]). of bone marrow cell population; and 3) No evidence of 27 28 Abroun et al is as yet unknown. B-cells originate from hematopoietic stem TABLE. Multiple Myeloma Staging System* cells (HSCs) in BM. Progenitor B-cells (B220+, CD19−, + International Staging CD117 ), B220 in mice, known as CD45 in humans, + + + + Stage System (ISS) Durie-Salmon and precursor B-cells (B220 , CD19 , CD25 , CD38 ) All of: Hb > 10 g/dL, normal calcium; are derived from HSCs and finally differentiated to mature + + + − β2-microglobulin skeletal survey: normal or single plas- B-cells (CD19 , CD20 , CD24 , CD93 ). Plasma cells + − − + + + + I (β2M) < 3.5 mg/L and macytoma or osteoporosis, serum (CD9 , CD19 CD24 , CD38 , FcγRII , CXCR4 , Blimp1 albumin ≥ 3.5 g/dL paraprotein level IgG < 5 g/dL, IgA < 3 [B-lymphocyte–induced maturation protein 1]) derived from g/dL, urinary light chain excretion activated B-cells [12-14]. Myeloma cells are the transformed β2M < 3.5 and albumin counterparts of post–germinal-center BM plasma blasts or II Fulfilling the criteria of neither I nor III < 3.5 plasma cells [15]. In silico tools offer an attractive alternative One or more of: Hb < 8.5 g/dL, high strategy to the cumbersome experimental approaches [16]. calcium > 12 mg/dL; skeletal survey: These computational tools have metamorphosed over the III β2M ≥ 5.5 mg/L three or more lytic bone lesions, years into complex algorithms that attempt to efficiently pre- serum paraprotein IgG > 7 g/dL,
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