尿标本量和沉渣制片存放条件对尿足细胞检测的影响

The Inflance of Podocytes Detection in Urine Sampie Quantities and Storage

目的:探讨少量尿液存放后检测尿足细胞的方法。方法:采用免疫荧光法鉴别足细胞的基础上,从尿标本量,沉渣制片存放温度、时间及一抗孵育时间与原检测方法比较,以足细胞阳性检出率、足细胞数量为指标评估实用性。结果:(1)尿量10ml与100ml(原方法)足细胞检出数相关性较好(r=0.670,P<0.05)。(2)沉渣制片标本于-20℃和4℃存放3d检出率分别为88%、63%,与原方法比较差异无统计学意义(P>0.05);存放7d阳性检出率分别为38%、38%,与原方法比较(P<0.05);同样存放3d对足细胞检出数影响不大,而7d足细胞检出数为0(0,9)、0(0,10.75)与原方法27.5(7.25,58.25)比较,差异有统计学意义(P<0.05);温度对同一时间点足细胞检出率和计数无明显影响。(3)一抗孵育1h与过夜(原方法)对足细胞检出率无影响,足细胞检出数分别为15(6,31)/20HP和8(3,17)/20HP,差异有统计学意义(P<0.05)。结论:采用免疫荧光法检测尿足细胞采尿标100ml,连续制片效果较好。临床上常遇肾衰竭少尿患者,可用10ml尿标本,沉渣制片存放4℃,3d内完成。

Objective：To investigate the improvement of method for the detection of urinary podocytes by immunofluorescence.Methods：The previous method for the detection of urinary podocytes by immunofluorescence was modified in the quantity of urine sample,storage temperature,duration of urine sedimentation slices and incubation time of first antibody.The method was evaluated by the quantity and detection rate of podocytes.Results：The quantity of podocytes in 10ml urine samples was well correlated to that in 100ml urine samples （r=0.670,P〈0.05）.The detection rates from urine sedimentation slices stored in -20 ℃ and 4 ℃ for 7 days were 38% and 38%,respectively,which were significantly lower than that in previous method （P〈0.05）.No difference was found for the storage in -20 ℃ and 4 ℃ for 3 days （P〉0.05）.However,when it came to 7 days,the podocytes detected were significantly lower than those with previous method（0[0, 9] vs 27.5[7.25,58.2],P〈0.05;0（0,10.75） vs 27.5（7.25,58.2）,P〈0.05,respectively.There was no significant difference in the detection rate of the incubation of first antibody for 1 hour and overnight,while the quantity of podocytes was significantly higher in the latter[15（6,31）/20 HP vs 8（3,17）/20 HP,P〈0.05].Conclusion：Enough samples should be provided for the detection of urinary podocytes by immunofluorescence.Ten ml sample could be used when patients with oliguria and acute renal failure,and the urine sedimentation slices could be kept in 4 ℃ for less than 3 days.