استانده كردن و اصلاح روش فلوئورومتري براي اندازه گيري هيستامين خون تام

مجله علمي دانشگاه علوم پزشكي و خدمات بهداشتي درماني همدان

دوره 3 - شماره 5

نوع مقاله: ---- Unspecified ----
چكيده:

هيستامين يكي از ميانجي هاي اصلي در اغلب بيماري هاي آلرژيك است و لذا اندازه گيري آن مي تواند در تشخيص اين قبيل بيماري ها بسيار ارزشمند باشد. اساسا سه روش براي اندازه گيري هيستامين خون وجود دارد كه عبارتند از: 1) پرتوآنزيمي Rradioenzymatic و 2) عيارگيري پرتوايمني RIA) Radioimmunoassay) و 3) فلوئورومتري. روش فلوئورومتري در مقايسه با دو روش ديگر داراي اين مزيت است كه در آن از مواد پرتوزا استفاده نمي شود. براي استانده كردن اندازه گيري هيستامين خون تام به روش فلوئورومتري لازم بود دقت و درستي آن تعيين شود. بدين منظور منحني استاندارد رسم و يك نمونه واحد بصورت 10 لوله اي در 10 روز متوالي آزمايش گرديد و ضريب تغييرات محاسبه شد (دقت). براي تعيين درستي از روش محاسبه درصد بازيافت با انجام آزمايش بر روي يك نمونه واحد به مدت 10 روز متوالي استفاده گرديد. با توجه به حجم نسبتا زياد محلول هاي مورد نياز و خون تام براي انجام آزمايش (5 ميلي ليتر براي هر لوله) روي فوق بر روي يك ميلي ليتر خون تام اصلاح شد. بدين منظور مجددا يك منحني استاندارد رسم و دقت و درستي روش بصورت پيش گفته تعيين و با روش اصلي مقايسه گرديد. نتايج حاصله دقت و درستي نسبتا بالاي روش اصلاح شده را نشان داد.

Standardization and modification of Fluorometric method of Histamine assaying in whole blood
Article Type: ---- Unspecified ----
Abstract:

Histamine (Hm), formed during decarboxylation of L-histidine by the action of L-histidine decarboxylase, a pyridoxal phosphate dependent enzyme, is one of the main mediators in such allergic diseases as asthma, urticaria and had fever. Hence, Hm assay can be used as a diagnostic tool in these disorders. There are three main methods for HM assaying: 1) Radio enzymatic, 2) Radioimmunoassay (RIA) and 3) Fluorometric. The latter method has the advantage of not using radioactive materials. In order to standardize fluorometric assay, the precision and accuracy of the method had to be determined. To do this, standard curve of Hm was drawn and then a single blood sample (Taken from IBTS) was tested in 10 tubes for 10 consecutive days. The range of result differences in 100 tests was 5 ng/ml and for average of results in 10 days was 1.4 ng/ml. Also, the coefficient of variation (C.V.) was 4.01% (Precision). Accuracy of the method was evaluated using recovery percent. This was also done on a single blood sample for 10 consecutive days. The range of recovery percent was 92-98%. Since in fluorometric assays it is preferable to do the tests in triplicate and considering the rather large volume of blood needed for Hm assaying fluorometrically in triplicate (15ml), it was intended to modify the original method so that only 1ml (3ml for triplicate) could be used for the test. To do this a new standard curve of Hm was drawn and precision and accuracy of the modified method was evaluated against the original one, in the same manner as mentioned above. The results of this study demonstrated the rather high precision and accuracy of the modified method.

قیمت : 20,000 ريال