Morales García A I, Górriz Teruel J L, Plancha Mansanet M C, Escudero Quesada V, Pallardó Mateu L M
Servicio de Nefrología, Hospital Universitario Dr. Peset, Valencia.
Nefrologia. 2009;29(4):331-5. doi: 10.3265/Nefrologia.2009.29.4.5377.en.full.
The measurement of i-PTH circulating is not easy due to its analytical variablity. Variability that appears in the process that goes from the sample collection to the final result determination. There are several important aspects that can influence within the pre-test variability: type of sample (serum o plasma), temperature, time elapses from blood extraction to freezing and from freezing to i-PTH quantification. Blood coming from centres far from our laboratory do not always meet the required processing conditions. Our aim was to study the stability of i-PTH with varying conditions of temperature and time until freezing in patients with chronic kidney disease (CKD).
We have analyzed 294 blood samples of 49 patients with chronic kidney disease (18 transplantated patients (36.7%) and 31 patients in haemodyalisis (63.3%)). The blood samples were collected using tubes treated with ethylenediaminotetraacetic acid (EDTA); these samples were subjected to different conditions of temperature and time before they were frozen, constituting 6 groups: blood centrifuged and plasma immediately frozen (group A or reference group); blood maintained 1 hour at room temperature and plasma stored at 2-8 masculineC during 0, 8 and 24 hours (groups B,C,D); blood maintained 3 hours at room temperature and plasma stored at 2-8 masculineC during 0 and 8 hours (groups E,F). The intact PTH (i-PTH) was measured using the immunoradiometric assay (IRMA Total Intact Scantibodies assay). We have analyzed the differences between the PTH-i mean values in the referenced groud and the others. We have applied the tests of homogeneity variance and normality and we have perform a comparation by pairs with the t-test including the Bonferroni correction.
The mean value of intact-PTH in the referente Group was 202.5+/-199.72 pg/ml. The means values of intact-PTH in the other groups were 196 +/- 203.23 pg/ml, 202.8 +/- 200.2 pg/ml, 200.06 +/- 194.87 pg/ml, 204.08 +/- 204.073 pg/ml, 197.94 +/- 182.31 pg/ml. The results were practically identical for each group. We did not find important differences with respect to the reference group (p = 0.87, p = 0,99, p = 0,95, p = 0,96, p = 0,90 when comparing with groups 2a, 2b, 2c, 3a y 3b).
The use of EDTA maintain the PTH stability during a longer period without the necessity of freezing the samples immediately. These results can help to state strategies to management the samples in patients with ERC.
由于其分析变异性,循环中全段甲状旁腺激素(i-PTH)的测量并不容易。这种变异性出现在从样本采集到最终结果测定的过程中。在检测前的变异性中有几个重要方面会产生影响:样本类型(血清或血浆)、温度、从血液抽取到冷冻以及从冷冻到i-PTH定量所经过的时间。来自距离我们实验室较远中心的血液并不总是满足所需的处理条件。我们的目的是研究慢性肾脏病(CKD)患者在不同温度和时间条件下直至冷冻时i-PTH的稳定性。
我们分析了49例慢性肾脏病患者的294份血样(18例移植患者(36.7%)和31例血液透析患者(63.3%))。使用经乙二胺四乙酸(EDTA)处理的试管采集血样;这些样本在冷冻前经历不同的温度和时间条件,构成6组:血液离心后血浆立即冷冻(A组或参照组);血液在室温下保存1小时,血浆在2 - 8℃保存0、8和24小时(B、C、D组);血液在室温下保存3小时,血浆在2 - 8℃保存0和8小时(E、F组)。使用免疫放射分析法(IRMA总全段甲状旁腺激素检测法)测量全段甲状旁腺激素(i-PTH)。我们分析了参照组与其他组中i-PTH均值之间的差异。我们应用了方差齐性检验和正态性检验,并通过配对t检验(包括Bonferroni校正)进行比较。
参照组中全段甲状旁腺激素的均值为202.5±199.72 pg/ml。其他组中全段甲状旁腺激素的均值分别为196±203.23 pg/ml、202.8±200.2 pg/ml、200.06±194.87 pg/ml、204.08±204.073 pg/ml、197.94±182.31 pg/ml。每组结果实际相同。与参照组相比,我们未发现显著差异(与2a、2b、2c、3a和3b组比较时p = 0.87、p = 0.99、p = 0.95、p = 0.96、p = 0.90)。
使用EDTA可在较长时间内维持PTH的稳定性,无需立即冷冻样本。这些结果有助于制定慢性肾脏病患者样本管理策略。