使用基于胸苷酸激酶的抗原和抗体捕获酶免疫测定法对HIV合并感染者中的活动性疾病进行血清学诊断。
Sero-diagnosis of Active Disease among HIV Co-infected Persons using Thymidylate Kinase based Antigen and Antibody Capture Enzyme Immuno-Assays.
作者信息
Wayengera Misaki, Mwebaza Ivan, Welishe Johnson, Nakimuli Cynthia, Kateete David P, Wampande Eddie, Kirimunda Samuel, Bayigga Lois, Musubika Carol, Babirye Peace, Asiimwe Benon, Joloba Moses L
机构信息
Department of Pathology, Unit of Genetics and Genomics, School of Biomedical Science, Makerere University College of Health Sciences, Kampala, Uganda.
Department of Immunology/Molecular Biology/Mycobacteriology, School of Biomedical Sciences, Makerere University College of Health Sciences, Uganda.
出版信息
Mycobact Dis. 2017 Jun;7(2). doi: 10.4172/2161-1068.1000241. Epub 2017 May 31.
BACKGROUND
Clinical and laboratory diagnosis of Active Tuberculosis (ATB) and latent () infections (LTBI) among people living with HIV/AIDS (PLWHA) presents formidable challenges. In the past, WHO issued an advisory against the use of existing TB sero-diagnostics. Emerging evidence, however, points to a precision of TB sero-diagnostics based on secretory rather than structural antigens. We hypothesized that secretory levels of thymidylate kinase (TMKmt) can Designate ATBI from LTBI and no TB (NTB). Here, we report in-house validation studies of levels of TMKmt antigen (Ag) and host specific TMKmt antibody (Ab) amongst HIV +ve and HIV -ve participants.
METHODS AND RESULTS
Direct TMKmt Ag and host specific IgG Ab detection EIAs were conducted on broadly consented, stored serum (N=281[Ag] . 214 [Ab] respective) samples stratified as either HIV +ve or HIV-ve ATB relative to LTBI and No TB. On one hand, UG-peptide 1 and its PAb-based EIAs accurately diagnosed ATB relative to LTBI and NTB among HIV +ve subjects {irrespectively: (a) Ag detection ATB=OD>0.490; 95% CI: 0.7446 to 0.8715 . LTBI=OD<0.490; 95% CI 0.4325 to 0.4829 . NTB=OD<0.26; 95% CI 0.1675 to 0.2567 and (b) TMKmt specific IgG detection ATB=OD>1.00; 95% CI 1.170 to 1.528 [HIV +ve] and 2.044 to 2.978 [HIV -ve] respectively . LTBI=OD<1.00; 95% CI 0.2690 to 0.6396 . NTB=OD<; 95% CI 0.1527 to 0.8751}. HIV -ve ATB presented with Ag levels greater than NTB and less than LTBI (i.e. ATB -ve=<0.490 ODs>0.26), but displayed better ant-TMKmt IgG responses (OD>2.00; 95% CI 2.044 to 2.978) relative to HIV +ve ATB (OD<1.600; 95% CI 1.170 to 1.528); suggesting a better control of -septicemia. On the other hand, UG-peptide 2 and its PAb-based EIAs did not demonstrate ATB diagnostic potential regardless of HIV sero-status, except towards designating NTB.
CONCLUSIONS
TMKmt Ab and Ag detecting EIAs based on UG-peptide 1 and its derivative PAb can accurately demarcate ATB from LTBI and NTB among HIV +ve subjects.
背景
对艾滋病毒/艾滋病感染者（PLWHA）中的活动性结核病（ATB）和潜伏性结核感染（LTBI）进行临床和实验室诊断面临巨大挑战。过去，世界卫生组织发布了一项反对使用现有结核病血清学诊断方法的建议。然而，新出现的证据表明，基于分泌性而非结构性抗原的结核病血清学诊断具有更高的准确性。我们假设胸苷酸激酶（TMKmt）的分泌水平可以区分活动性结核感染（ATBI）与潜伏性结核感染（LTBI）及非结核（NTB）。在此，我们报告了在HIV阳性和HIV阴性参与者中对TMKmt抗原（Ag）水平和宿主特异性TMKmt抗体（Ab）进行的内部验证研究。
方法与结果
对广泛同意参与研究的储存血清样本（分别为N = 281个[Ag]样本、214个[Ab]样本）进行直接TMKmt Ag和宿主特异性IgG Ab检测酶联免疫吸附测定（EIA），这些样本根据HIV阳性或HIV阴性、相对于LTBI和非结核的ATB进行分层。一方面，UG肽1及其基于多克隆抗体（PAb）的EIA在HIV阳性受试者中能够准确诊断ATB与LTBI和NTB（分别为：（a）Ag检测：ATB = OD>0.490；95%置信区间：0.7446至0.8715，LTBI = OD<0.490；95%置信区间0.4325至0.4829，NTB = OD<0.26；95%置信区间0.1675至0.2567；（b）TMKmt特异性IgG检测：ATB = OD>1.00；95%置信区间分别为1.170至1.528[HIV阳性]和2.044至2.978[HIV阴性]，LTBI = OD<1.00；95%置信区间0.2690至0.6396，NTB = OD<；95%置信区间0.1527至0.8751）。HIV阴性的ATB患者的Ag水平高于NTB且低于LTBI（即ATB阴性=<0.490 ODs>0.26），但相对于HIV阳性的ATB患者（OD<1.600；95%置信区间1.170至1.528），其抗TMKmt IgG反应更好（OD>2.00；95%置信区间2.044至2.978）；这表明对败血症有更好的控制。另一方面，UG肽2及其基于PAb的EIA无论HIV血清学状态如何，除了用于区分NTB外，均未显示出ATB诊断潜力。
结论
基于UG肽1及其衍生PAb的TMKmt Ab和Ag检测EIA能够在HIV阳性受试者中准确区分ATB与LTBI和NTB。
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