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Thread: Platelets, Normal EDTA anticoagulated blood picture - blood histology atlas

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    Default Platelets, Normal EDTA anticoagulated blood picture - blood histology atlas

    Pseudothrombocytopenia is usually associated with blood specimens anticoagulated with EDTA or other anticoagulants. EDTA salt is an anticoagulant frequently used in automated blood cell counters, which can maintain cell formation and prevent platelets from aggregating. The use of this anticoagulant is known to cause erroneous reports of low platelet counts by automated analyzers. In automated cell counting of K2-EDTA anticoagulated blood samples, thrombocytopenia is occasionally observed that has no relation to any underlying disease. This phenomenon of EDTA-induced pseudothrombocytopenia may occur in healthy people and in patients with a variety of diseases when their blood samples are anticoagulated with EDTA.2 The prevalence rate of EDTA-induced pseudothrombocytopenia was reported to be 0.07% to 0.20%. For hospitalized patients, an incidence of 0.1% to 2.0% was reported. Up to 17% of the patients referred to the outpatient clinic for isolated thrombocytopenia were found to have EDTA-induced pseudothrombocytopenia.

    The phenomenon of EDTA-induced pseudothrombocytopenia caused by a variety of factors has been reported in recent years.10 Pseudothrombocytopenia is a spuriously low platelet count caused by agglutinating antibodies that induce in vitro platelet clumping. Most of the agglutinins react most strongly at room temperature or lower temperatures, but some are temperature-independent or react best at 37°C.11 Most are IgG, but IgM, IgA, and combinations of IgG and IgM or IgA and IgG have also been described.12 The observation that platelets from patients with Glanzmann disease do not react with these antibodies suggests that platelet glycoprotein (GP) IIb/IIIa is involved in EDTA-induced antibody reactions.13,14 This hypothesis is further supported by the identification of a cold antibody (IgM) directed against platelet GPIIb that causes pseudothrombocytopenia.

    The failure to recognize this form of EDTA-induced pseudothrombocytopenia may lead to a person with a normal platelet count being considered as having severe thrombocytopenia.15 Unnecessary evaluations, canceled surgical procedures, avoidance of conduction anesthesia, unwarranted splenectomies, and undue patient expense and anxiety are potential outcomes for person with this form of in vitro artifact. Since its initial description in 1969, this problem has become commonly associated with hospitalized patients, especially seriously ill patients. EDTA-induced pseudothrombocytopenia has also been reported, although less commonly, in healthy people undergoing routine blood cell counts.

    Pseudothrombocytopenia is a clinical problem that may lead to misdiagnosis, unnecessary examination, and inappropriate treatment. Therefore, to establish a proper method to prevent inaccurate platelet counting is very important. It has been reported that the platelet aggregation by EDTA could be prevented by supplementing with aminoglycosides.4 Jeon and Yang16 showed that kanamycin may prevent and dissociate platelets from aggregation in a child with EDTA-induced pseudothrombocytopenia. In the present study, we investigated a rare case of multianticoagulant-dependent pseudothrombocytopenia in a 35-year-old man, for whom the inhibition of multianticoagulant-dependent pseudothrombocytopenia was available only with amikacin but not with vitamin B6, aminophylline, or gentamicin. To our knowledge, this is the first report on the inhibition and dissociation effects of amikacin on multianticoagulant-dependent pseudothrombocytopenia.

    Materials and Methods
    Vacuum blood tubes containing K2-EDTA, heparin lithium, 109 mmol/L of sodium citrate, or sodium fluoride (NaF) were purchased from Guangzhou Improve Medical Instruments, Guangzhou, China. Amikacin, aminophylline, vitamin B6, and gentamicin were purchased from Qilu Pharmaceutical, Jining, China; Guangzhou Ming Xing Pharmaceutical, Guangzhou, China; Guangzhou Baiyunshan Pharmaceutical, Guangzhou, China; and Shandong Lukang Cisen Pharmaceutical, Jining, China. Blood cells were counted by using a CD-3700 blood cell counter (Abbott Laboratories, Abbott Park, IL) at room temperature at regular intervals after sampling. Reagents and quality control substances were provided by Abbott Laboratories.
    Platelets, Normal EDTA anticoagulated blood attachment.php?s=cb017e5c967c04a7d481f13d4e24f43f&attachmentid=292&d=1435539129

    A 35-year-old man, a healthy surgeon, had a WBC count of 6,500/μL (6.5 × 109/L). His platelet count (56–78 × 103/μL [56–78 × 109/L]) was lower than the reference range according to an electronic counting method, and the phenomenon had lasted for more than 2 years. The patient stated he had not had epistaxis, melena, hematuria, easy bruising, or oral mucosal bleeding. A physical examination showed no petechiae or bleeding. Platelet counts obtained by a CD-3700 blood cell counter were significantly different from those obtained by a manual method (mean ± SD, 210 ± 8 × 109/L), and peripheral blood smears proved that the platelet form and the platelet count were normal. K2-EDTA anticoagulant resulted in spuriously low platelet counts, and blood samples anticoagulated with sodium citrate, NaF, or heparin lithium also showed an underestimation of the platelet count. Microscopic examination of the anticoagulated blood smears found adequate numbers of platelets, often in large clumps. These results, taken together with the clinical history, suggest that the patient had multianticoagulant-dependent pseudothrombocytopenia.

    The manual counting of platelets was completed in 15 minutes after blood sampling following the National Clinical Laboratory Operation Procedure and the manual result was used as the reference value. In blood samples anticoagulated with K2-EDTA, 109 mmol/L of sodium citrate, NaF, or heparin lithium, the platelet counts at different times (10 minutes and 0.5, , and 4 hours from the start point) were measured by the CD-3700 blood cell counter, and smear slides were obtained with manual method. Blood smears were prepared to observe the formation and distribution of platelets at the above mentioned times.

    The Effects of Auxiliary Reagents Before Treatment on CBCs The blood samples were previously supplemented separately with the auxiliary reagents amikacin (5 mg/mL of blood), aminophylline (6.5 mg/mL of blood), vitamin B6 (0.25 mg/mL of blood), or gentamicin (4,000 IU/mL of blood) and then mixed with K2-EDTA to observe changes of platelets at different times, so as to find the optimal auxiliary reagent. The optimal auxiliary reagent was added to the blood samples 10 minutes before or after sampling mixed with K2-EDTA, 109 mmol/L of sodium citrate, NaF, or heparin lithium, so as to observe its ability to inhibit and dissociate platelet aggregation.
    The Effects of Amikacin Supplementation on EDTA-Anticoagulated Blood Samples Amikacin was added to the EDTA-anticoagulated blood samples at 30, 60, 90, and 120 minutes after blood sample withdrawal, and platelets of control samples (untreated) and patient samples were counted after incubation at room temperature. Routine blood counts and the examination of blood cell morphologic features were performed in 30- to 60-minute intervals during the course of the observation.
    Amikacin was added to the blood samples with K2-EDTA to investigate the aggregated platelet dissociation using the platelet count instrument and microscopic examination during different times after drawing blood samples from the patient with multianticoagulant-dependent pseudothrombocytopenia and a 50-year-old woman with intrinsic thrombocytopenia accompanying EDTA-induced pseudothrombocytopenia.

    Data are expressed as mean ± SD. The Student paired t test was used for statistical comparisons. A P value of less than .05 was regarded as significant.

    References:
    Amikacin Can Be Added to Blood to Reduce the Fall in Platelet Count











    Last edited by Medical Photos; 08-14-2015 at 10:28 PM.

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    Default Platelets Normal EDTA anticoagulated blood picture blood histology atlas

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    Really nice post. I like it.

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