Rapid, novel, specific, high-throughput assay for diagnosis of Loa loa infection
Rapid, novel, specific, high-throughput assay for diagnosis of Loa loa infection
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DOI：
10.1128/jcm.00490-08
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发表时间：
2008-07-01
影响因子：
9.4
通讯作者：
Nutman, Thomas B.
中科院分区：
文献类型：
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作者：
Burbelo, Peter D.;Ramanathan, Roshan;Nutman, Thomas B.
轻松准确地诊断LOA感染的能力仍然是一项艰巨的任务。在现有的诊断方法中，许多方法不适用于现场护理点测试。为了探讨荧光素酶免疫沉淀系统(LIPS)能否用于快速、特异地诊断Lloa感染，建立了一种基于重组Lloa SXP-1(命名为LlSXP-1)抗原的免疫球蛋白G(Ig G)和Ig G4亚类抗体的LIPs检测方法，并用健康对照或已证实感染了Lloa、曼氏曼氏杆菌、螺旋体、珊瑚类圆线虫或班氏吴氏杆菌的患者的血清进行了检测。与基于LlSXP-1 IgG4的酶联免疫吸附试验(67%的灵敏度和99%的特异度)相比，嘴唇试验检测LlSXP-1容易区分来自未感染患者的L.LoA感染的抗体的敏感性和特异性显著提高。未观察到明显的免疫反应，但少数感染螺旋体、Perstans或Wancrofti的患者免疫反应阳性。检测抗L1SXP-1的特异性抗体与抗-Ig G结果有显著相关性(r=0.85P<；0.00001)，但与单独检测总的Ig G反应无明显差异。相比之下，快速LIPS格式(称为QLIPS)在非平衡条件下在不到15分钟的时间内进行测试，显著提高了交叉反应旋毛虫患者血清的特异性(100%敏感性和100%特异性)。这些结果表明，LIPS(以及更快速的QLIPS测试)代表着诊断L.LoA感染能力的重大进步，并可能在未来的护理点诊断中应用。
The ability to diagnose Loa loa infection readily and accurately remains a demanding task. Among the available diagnostic methods, many are impractical for point-of-care field testing. To investigate whether luciferase immunoprecipitation systems (LIPS) can be used for rapid and specific diagnosis of L. loa infection, a LIPS assay was developed based on immunoglobulin G (IgG) and IgG4 subclass antibodies to a recombinant L. loa SXP-1 (designated LlSXP-1) antigen and tested with sera from healthy controls or patients with proven infection with L. loa, Mansonella perstans, Onchocerca volvulus, Strongyloides stercoralis, or Wuchereria bancrofti. A LIPS test measuring IgG antibody against LlSXP-1 readily differentiated L. loa-infected from uninfected patients and demonstrated markedly improved sensitivity and specificity compared with an LlSXP-1 IgG4-based enzyme-linked immunosorbent assay (67% sensitivity and 99% specificity). No significant immunoreactivity was observed with S. stercoralis-infected sera, but a small number of patients infected with O. volvulus, M. perstans, or W. bancrofti showed positive immunoreactivity. Measuring anti-IgG4-specific antibodies to LlSXP-1 showed a significant correlation (r similar to 0.85; P < 0.00001) with the anti-IgG results but showed no advantage over measuring the total IgG response alone. In contrast, a rapid LIPS format (called QLIPS) in which the tests are performed in less than 15 minutes under nonequilibrium conditions significantly improved the specificity for cross-reactive O. volvulus patient sera (100% sensitivity and 100% specificity). These results suggest that LIPS (and the even more rapid test QLIPS) represents a major advance in the ability to diagnose L. loa infection and may have future applications for point-of-care diagnostics.