改良蝰蛇毒磷脂时间试验和硅凝固时间试验检测狼疮抗凝物的性能验证

doi:10.3969/j.issn.1673-8640.2017.03.016

摘要/Abstract

摘要：

目的对基于2种不同原理的狼疮抗凝物（LA）检测方法,即改良蝰蛇毒磷脂时间（dRVVT）试验和硅凝固时间（SCT）试验进行全面的方法学评估,以期为临床实验室选择LA检测方法提供参考依据。方法计数50例乏血小板血浆（PPP）血小板（PLT）数量,验证现有离心条件是否满足要求。按照美国临床实验室标准化协会文件及我国《临床血液学检验常规项目分析质量要求》（WS/T406-2012）文件,对2种LA检测方法的精密度、携带污染率、方法学比对进行全面评估,并建立了不同方法检测LA的参考范围（RI）/临界值（Cut-off）。结果 50例PPP的PLT计数为（0~9）×10⁹/L,满足指南<10×10⁹/L的要求。dRVVT试验和SCT试验3个结果表达方式的批内精密度均<2.50%。dRVVT试验批间精密度为2.76%~3.53%,SCT试验批间精密度为6.23%~7.02%。标准化比值（NR）的批间精密度<其他2种结果表达方式。所有检测方法携带污染率绝对值<3.00%。dRVVT NR计算所得RI/Cut-off值为1.24,高于试剂说明书数据（1.20）;SCT NR计算所得RI/Cut-off值为1.16,与说明书中一致。不同仪器同种方法和相同仪器不同方法之间比对数据相关性良好（r>0.900）。同一仪器2种检测方法NR之间（P=0.248）和2种方法阴阳性判断之间差异均无统计学意义（P=0.068）。结论基于不同原理的dRVVT试验和SCT试验均有良好的检测性能,各实验室建立或验证了适合本实验室的RI/Cut-off值后,适用于LA的临床常规检测。

关键词: 狼疮抗凝物, 改良蝰蛇毒磷脂时间试验, 硅凝固时间试验, 性能验证

Abstract:

Objective To evaluate 2 lupus anticoagulant （LA） assays based on diluted Russell viper venom time （dRVVT） and silica clotting time （SCT）,and to provide a reference to choose suitable assay for LA determination. Methods The platelet （PLT） counts of 50 cases of poor platelet plasma（PPP） were counted to validate centrifuge procedure. According to the Clinical and Laboratory Standards Institute （CLSI）documents and the Analytical Quality Specifications for Routine Tests in Clinical Hematology（WS/T406-2012）,their precisions,carryover rates and methodological comparisons were evaluated. The reference interval （RI）/cut-off value was also established. Results PLT count was （0-9）×10⁹/L in the 50 cases of PPP,and met the requirements of guidelines （<10×10⁹/L）. The within-run precisions of dRVVT and SCT were <2.50%. The between-run precisions of dRVVT and SCT were 2.76%-3.53% and 6.23%-7.02%,respectively. The precision of normalized ratio was < the other 2 expressions. The absolute values of carryover rates of the 2 assays were < 3.00%. The RI/cut-off value of dRVVT was 1.24,which was higher than that provided by manufactures' instructions （1.20）. The RI/cut-off value of SCT was 1.16,which was as the same as that provided by manufactures' instructions. The r of the same assay in different instruments and different assays in the same instrument were >0.900. There was no statistical significance for NR between the 2 assays in the same instrument（P=0.248）,and there was no statistical significance between 2 methods in the evaluation of positive and negative results. Conclusions The 2 LA assays based on dRVVT and SCT have good performance. Every laboratory should establish local LA RI/cut-off values.

Key words: Lupus anticoagulant, Diluted Russell viper venom time, Silica clotting time, Performance validation

中图分类号:

R446.1

寿玮龄, 陈倩, 吴卫, 范连凯, 崔巍. 改良蝰蛇毒磷脂时间试验和硅凝固时间试验检测狼疮抗凝物的性能验证[J]. 检验医学, 2017, 32(3): 224-228.

SHOU Weiling, CHEN Qian, WU Wei, FAN Liankai, CUI Wei. Performance of lupus anticoagulant assays based on diluted Russell viper venom time and silica clotting time[J]. Laboratory Medicine, 2017, 32(3): 224-228.

图/表 4

参考文献 21

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序号	全血	PPP	序号	全血	PPP	序号	全血	PPP	序号	全血	PPP
1	287	8	14	192	6	27	246	1	40	80	4
2	16	0	15	754	1	28	186	0	41	74	3
3	117	2	16	636	1	29	203	1	42	27	2
4	164	0	17	353	0	30	240	1	43	19	1
5	226	7	18	431	0	31	160	1	44	65	1
6	539	3	19	482	0	32	281	5	45	147	2
7	584	1	20	386	1	33	297	0	46	158	4
8	603	1	21	449	2	34	95	1	47	310	9
9	960	2	22	223	1	35	180	1	48	376	8
10	799	5	23	323	1	36	46	0	49	379	5
11	1 114	7	24	551	7	37	657	5	50	540	7
12	461	8	25	407	4	38	411	5
13	342	5	26	311	3	39	461	3

序号	全血	PPP	序号	全血	PPP	序号	全血	PPP	序号	全血	PPP
1	287	8	14	192	6	27	246	1	40	80	4
2	16	0	15	754	1	28	186	0	41	74	3
3	117	2	16	636	1	29	203	1	42	27	2
4	164	0	17	353	0	30	240	1	43	19	1
5	226	7	18	431	0	31	160	1	44	65	1
6	539	3	19	482	0	32	281	5	45	147	2
7	584	1	20	386	1	33	297	0	46	158	4
8	603	1	21	449	2	34	95	1	47	310	9
9	960	2	22	223	1	35	180	1	48	376	8
10	799	5	23	323	1	36	46	0	49	379	5
11	1 114	7	24	551	7	37	657	5	50	540	7
12	461	8	25	407	4	38	411	5
13	342	5	26	311	3	39	461	3

试验	批内精密度		批间精密度
试验	水平1[CV（%）]	水平2[CV（%）]	水平1[CV（%）]	水平2[CV（%）]
dRVVT
凝固时间	30.26±0.42（1.40）	84.65±0.67（0.79）	36.54±0.88（2.41）	66.12±2.33（3.53）
R	1.13±0.01（1.22）	2.37±0.02（0.98）	1.12±0.03（3.02）	1.88±0.06（2.97）
NR	1.00±0.01（1.05）	2.08±0.02（0.92）	0.99±0.02（2.37）	1.66±0.05（2.76）
SCT
凝固时间	38.54±1.58（1.49）	116.40±1.35（1.16）	33.51±1.61（4.80）	75.19±5.28（7.02）
R	1.13±0.03（2.35）	3.31±0.06（1.77）	0.91±0.04（3.94）	1.90±0.13（6.64）
NR	1.08±0.18（1.64）	3.19±0.06（1.77）	0.88±0.03（3.64）	1.84±0.11（6.23）

试验	批内精密度		批间精密度
试验	水平1[CV（%）]	水平2[CV（%）]	水平1[CV（%）]	水平2[CV（%）]
dRVVT
凝固时间	30.26±0.42（1.40）	84.65±0.67（0.79）	36.54±0.88（2.41）	66.12±2.33（3.53）
R	1.13±0.01（1.22）	2.37±0.02（0.98）	1.12±0.03（3.02）	1.88±0.06（2.97）
NR	1.00±0.01（1.05）	2.08±0.02（0.92）	0.99±0.02（2.37）	1.66±0.05（2.76）
SCT
凝固时间	38.54±1.58（1.49）	116.40±1.35（1.16）	33.51±1.61（4.80）	75.19±5.28（7.02）
R	1.13±0.03（2.35）	3.31±0.06（1.77）	0.91±0.04（3.94）	1.90±0.13（6.64）
NR	1.08±0.18（1.64）	3.19±0.06（1.77）	0.88±0.03（3.64）	1.84±0.11（6.23）

仪器	例数	dRVVT			SCT
仪器	例数	凝固时间	R	NR	凝固时间	R	NR
仪器1	30	44.70±15.70	1.42±0.50	1.25±0.33	62.73±44.15	1.63±1.15	1.18±0.62
仪器2	30	46.40±15.95	1.50±0.50	1.30±0.95	53.10±31.20	1.50±0.90	1.18±0.52
平均百分偏差（%）		4.44	6.35	2.10	-11.29	-2.88	8.59
r值		0.963	0.954	0.954	0.964	0.915	0.915
t值		2.091	2.269	1.298	-3.215	-2.302	0.191
P值		0.045	0.031	0.204	0.003	0.029	0.850