检测尿Ⅳ型胶原的磁微粒化学发光法的建立及尿Ⅳ型胶原在膜性肾病中的临床应用

doi:10.3969/j.issn.1673-8640.2018.05.016

摘要/Abstract

摘要：

目的建立检测尿Ⅳ型胶原（ⅣC）的磁微粒化学发光法,并探讨尿ⅣC在膜性肾病（MN）中的临床应用价值。方法制备适用于尿ⅣC检测的标准品,建立检测尿ⅣC的磁微粒化学发光法,并对该方法的检测性能（检测限、线性范围、正确度、精密度、回收率及分析灵敏度）进行确认。采用受试者工作特征（ROC）曲线评估尿ⅣC对MN的诊断效能。采用该方法检测274例慢性肾病（CKD）患者及83名体检健康者（正常对照组）尿ⅣC水平,分析不同病理类型及不同分期的CKD患者尿ⅣC水平的变化。结果磁微粒化学发光法检测尿ⅣC的标准曲线拟合方程为Y=42.802X²+10 626X-10 856（r²=0.999）;最低检测限为1.48 ng/mL。低值、中值和高值样本的批内变异系数（CV）分别为4.22%、8.16%和3.46%,低值和中值批间CV分别为18.23%、17.61%,平均回收率为97.0%。ROC曲线分析显示,尿ⅣC诊断MN的曲线下面积（AUC）为0.875[95%可信区间（CI） 0.837~0.913],最佳临界值为4.15 ng/mL,敏感性为79.0%,特异性为86.7%。MN组、系膜增殖性肾小球肾炎（MsPGN）组、非膜性肾病（N-MN）组尿ⅣC水平均明显高于正常对照组（P<0.01）,MN组、MsPGN组尿ⅣC水平均明显高于N-MN组（P<0.01）。将MN患者根据CDK分期分为5期,CKD1期与CKD2、CKD3、CKD4期比较,尿ⅣC水平差异均有统计学意义（P<0.01）,而与CKD5期比较差异无统计学意义（P>0.05）。结论建立的磁微粒化学发光法操作简便、性能稳定可靠且价格低廉,适宜在临床推广应用,尿ⅣC对MN的诊断及预后判断具有较高的临床应用价值。

关键词: Ⅳ型胶原;, 尿液, 磁微粒化学发光法, 性能评价, 膜性肾病, 系膜增殖性肾小球肾炎

Abstract:

Objective To establish a magnetic particle chemiluminescence for the determination of urinary type Ⅳ collagen（ⅣC）,and to investigate the role of urinary ⅣC in membranous nephropathy（MN）. Methods A magnetic particle chemiluminescence was established for the determination of urinary ⅣC. The performance （determination limit,linear range,accuracy,precision,recovery rate and sensitivity） was verified. The receiver operating characteristic （ROC）curve was used to evaluate the efficiency of urinary ⅣC in the diagnosis of MN. The urinary ⅣC levels of 274 patients with chronic kidney disease（CKD） was determined,and the changes of urinary ⅣC levels in different pathological types and different CKD stages were compared. Results Urinary ⅣC was determined by magnetic particle chemiluminescence,and the standard curve was drawn,with the equation of Y= 42.802X²+10 626X-10 856 （r²=0.999）. The lower determination limit was 1.48 ng/mL,the within-run coefficients of variation（CV） for low-level, middle-level and high-level samples were 4.22%,8.16% and 3.46%,and the between-run CV for low-level and middle-level samples were 18.23% and 17.61%. The average recovery rate was 97.0%. ROC curve showed that the area under curve （AUC） of urinary ⅣC in the diagnosis of MN was 0.875 [95% confidence interval （CI）0.837-0.913]. The optimal cut-off value of urinary ⅣC was 4.15 ng/mL. The sensitivity was 79.0%,and the specificity was 86.7%. Compared with healthy control group,the levels of urinary ⅣC in MN group,mesangial proliferative glomerulonephritis（MsPGN） group and non-membranous nephropathy （N-MN）group were increased（P<0.01）. Compared with N-MN group,the levels of urinary ⅣC in MN group and MsPGN group were increased （P<0.01）. For CDK staging,there was a statistical significance in urinary ⅣC level between CKD1 group and CKD2,CDK3,CDK4 groups（P<0.01）. There was no statistical significance between CKD1 and CKD5 groups （P>0.05）. Conclusions Magnetic particle chemiluminescence for the determination of urinary ⅣC is simple,reliable and inexpensive,and it plays a role for the diagnosis and prognosis of MN.

Key words: Type Ⅳ collagen;, Urine, Magnetic particle chemiluminescence, Performance evaluation, Membranous nephropathy, Mesangial proliferative glomerulonephritis

中图分类号:

R446.1

王蕾, 陈伟琴, 徐旻一, 陈旭东. 检测尿Ⅳ型胶原的磁微粒化学发光法的建立及尿Ⅳ型胶原在膜性肾病中的临床应用[J]. 检验医学, 2018, 33(5): 442-446.

WANG Lei, CHEN Weiqin, XU Minyi, CHEN Xudong. Establishment of magnetic particle chemiluminescence for the determination of urinary type Ⅳ collagen and its clinical application in membranous nephropathy[J]. Laboratory Medicine, 2018, 33(5): 442-446.

图/表 6

参考文献 16

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稀释比例	理论值	实测值1	实测值2	实测均值
H	164.9	189.1	178.7	183.9
1H+1L	82.5	82.3	80.9	81.6
1H+3L	41.2	57.3	37.4	47.4
1H+7L	20.6	18.0	24.4	21.2
1H+15L	10.3	15.1	10.1	12.6
1H+31L	5.2	9.0	9.3	9.2

稀释比例	理论值	实测值1	实测值2	实测均值
H	164.9	189.1	178.7	183.9
1H+1L	82.5	82.3	80.9	81.6
1H+3L	41.2	57.3	37.4	47.4
1H+7L	20.6	18.0	24.4	21.2
1H+15L	10.3	15.1	10.1	12.6
1H+31L	5.2	9.0	9.3	9.2

样本号	理论值（ng/mL）	实测均值（ng/mL）	回收率（%）
1	15.8	15.1	95.8
2	54.3	53.2	98.0
3	77.5	75.4	97.3

样本号	理论值（ng/mL）	实测均值（ng/mL）	回收率（%）
1	15.8	15.1	95.8
2	54.3	53.2	98.0
3	77.5	75.4	97.3

CKD分期	例数	尿ⅣC
CKD1期	48	6.0（3.9~9.2）
CKD2期	36	13.6（7.9~24.8）^*
CKD3期	28	48.5（23.1~72.0）^*
CKD4期	11	33.0（18.0~97.9）^*
CKD5期	16	3.6（2.7~12.5）