A dried blood spot test for diagnosis of autoimmune pulmonary alveolar proteinosis.
A dried blood spot test for diagnosis of autoimmune pulmonary alveolar proteinosis.
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DOI：
10.1016/j.jim.2022.113366
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发表时间：
2022-12
影响因子：
2.2
通讯作者：
Trapnell, Bruce C.
中科院分区：
文献类型：
--
作者：
Carey, Brenna;Chalk, Claudia;Stock, Jennifer;Toth, Andrea;Klingler, Maria;Greenberg, Henry;Uchida, Kanji;Arumugam, Paritha;Trapnell, Bruce C.
关键词：
粒细胞/巨噬细胞集落刺激因子自身抗体（GMAb）介导自身免疫性肺泡蛋白沉积症（自身免疫性PAP）的发病机制，并且通过酶联免疫吸附测定（ELISA）-血清GMAb测试-在血清中定量它们是诊断自身免疫性PAP的“金标准”。由于GMAb在自身免疫PAP中较高，在健康人群中较低或无法检测，因此我们假设ELISA可适用于使用干血斑卡（DBSC）采集样本评价从指尖获得的血液。在这里，我们报告这样一种方法的开发-DBSC GMAb测试-并评估其测量血液中GMAb浓度和诊断自身免疫性PAP的能力。从60名自身免疫性PAP患者和19名健康人获得新鲜肝素化全血，并用于测量血液中的GMAb浓度（通过DBSC GMAb试验）。优化后，评价了DBSC GMAb检测的准确度、精密度、可靠性、灵敏度、特异性和耐用性。在孔间、板间、日间和操作员间变异方面，重复测量的变异系数较低，并且制备的DBSC标本暴露于宽范围的温度（−80°C至65°C）、重复冻融循环或在测试前储存长达2.5个月均不影响结果。血液中GMAb的空白限（LoB）、检测限（LoD）和定量下限（LLoQ）分别为0.01、0.21和3.5 μg/ml。受试者工作曲线特征分析确定2.7 μg/ml为区分自身免疫性PAP与健康人的最佳GMAb浓度截止值。该临界值低于LLoQ，自身免疫性PAP患者和健康人的GMAb结果范围广泛分离（中位数（四分位距）分别为：22.6（13.3 - 43.8）和0.23（0.20 - 0.30）μg/ml）。因此，建议将LLoQ作为指示阳性检测结果的范围下限（即，存在自身免疫PAP）;较低的值表示阴性测试结果（即，自身免疫性PAP不存在）。在30例自身免疫性PAP患者和19例健康人中，DBSC GMAb试验诊断自身免疫性PAP的敏感性和特异性均为100%。结果表明，DBSC GMAb测试可靠地测量血液中的GMAb，并且在自身免疫性PAP的诊断中表现良好。
Granulocyte/macrophage colony-stimulating factor autoantibodies (GMAbs) mediate the pathogenesis of autoimmune pulmonary alveolar proteinosis (autoimmune PAP) and their quantification in serum by enzyme-linked immunosorbent assay (ELISA) – the serum GMAb test – is the ‘gold standard’ for diagnosis of autoimmune PAP. Because GMAbs are high in autoimmune PAP and low or undetectable in healthy people, we hypothesized that the ELISA could be adapted for evaluation of blood obtained from the fingertip using a dried blood spot card (DBSC) for specimen collection. Here, we report development of such a method – the DBSC GMAb test – and evaluate its ability to measure GMAb concentration in blood and to diagnose autoimmune PAP. Fresh, heparinized whole blood was obtained from 60 autoimmune PAP patients and 19 healthy people and used to measure the GMAb concentration in blood (by the DBSC GMAb test). After optimization, the DBSC GMAb test was evaluated for accuracy, precision, reliability, sensitivity, specificity, and ruggedness. The coefficient of variation among repeated measurements was low with regard to well-to-well, plate-to-plate, day-to-day, and inter-operator variation, and results were unaffected by exposure of prepared DBSC specimens to a wide range of temperatures (from −80°C to 65°C), repeated freeze-thaw cycles, or storage for up to 2.5 months before testing. The limit of blank (LoB), limit of detection (LoD), and lower limit of quantification (LLoQ), were 0.01, 0.21, and 3.5 μg/ml of GMAb in the blood, respectively. Receiver operating curve characteristic analysis identified 2.7 µg/ml as the optimal GMAb concentration cutoff value to distinguish autoimmune PAP from healthy people. This cutoff value was less than the LLoQ and the ranges of GMAb results for autoimmune PAP patients and healthy people were widely separated (median (interquartile range): 22.6 (13.3 – 43.8) and 0.23 (0.20 – 0.30) μg/ml, respectively). Consequently, the LLoQ is recommended as the lower limit of the range indicating a positive test result (i.e., that autoimmune PAP is present); lower values indicate a negative test result (i.e., autoimmune PAP is not present). Among the 30 autoimmune PAP patients and 19 healthy people evaluated, the sensitivity and specificity of the DBSC GMAb test were both 100% for a diagnosis of autoimmune PAP. Results demonstrate the DBSC GMAb test reliably measures GMAbs in blood and performs well in the diagnosis of autoimmune PAP.
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DOI：
10.1164/ajrccm.162.2.9910032
发表时间：
2000-08-01
影响因子：
24.7
作者：
Kitamura, T;Uchida, K;Nakata, K
通讯作者：
Nakata, K
影响因子：
3.7
作者：
Smit PW;Sollis KA;Fiscus S;Ford N;Vitoria M;Essajee S;Barnett D;Cheng B;Crowe SM;Denny T;Landay A;Stevens W;Habiyambere V;Perriens JH;Peeling RW
通讯作者：
Peeling RW
影响因子：
11.1
作者：
Dube, Queen;Dow, Anna;Van Rie, Annelies
通讯作者：
Van Rie, Annelies
影响因子：
4.2
作者：
Mei, JV;Alexander, JR;Hannon, WH
通讯作者：
Hannon, WH
DOI：
10.1016/s0035-9203(99)90177-3
发表时间：
1999-01-01
影响因子：
2.2
作者：
Färnert, A;Arez, AP;do Rosário, V
通讯作者：
do Rosário, V