Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, United States of America.
The Peter Doherty Institute for Infection and Immunity, University of Melbourne & The Royal Melbourne Hospital, Melbourne, Victoria, Australia.
PLoS Negl Trop Dis. 2022 May 26;16(5):e0010442. doi: 10.1371/journal.pntd.0010442. eCollection 2022 May.
Antigen tests for diagnosis and disease monitoring in some types of neurocysticercosis (NCC) are useful but access to testing has been limited by availability of proprietary reagents and/or kits.
METHODS/PRINCIPAL FINDINGS: Three previously identified IgM-secreting hybridomas whose IgM products demonstrated specificity to Taenia solium underwent variable heavy and light chain sequencing and isotype conversion to mouse IgG. Screening of these recombinantly expressed IgG anti-Ts hybridomas, identified one (TsG10) with the highest affinity to crude Taenia antigen. TsG10 was then used as a capture antibody in a sandwich antigen detection immunoassay in combination with either a high titer polyclonal anti-Ts antibody or with biotinylated TsG10 (termed TsG10*bt). Using serum, plasma, and CSF samples from patients with active NCC and those from NCC-uninfected patients, ROC curve analyses demonstrated that the TsG10-TsG10-*bt assay achieved a 98% sensitivity and 100% specificity in detecting samples known to be antigen positive and outperformed the polyclonal based assay (sensitivity of 93% with 100% specificity). By comparing levels of Ts antigen (Ag) in paired CSF (n = 10) or plasma/serum (n = 19) samples from well-characterized patients with extra-parenchymal NCC early in infection and at the time of definitive cure, all but 2 (1 from CSF and 1 from plasma) became undetectable. There was a high degree of correlation (r = 0.98) between the Ag levels detected by this new assay and levels found by a commercial assay. Pilot studies indicate that this antigen can be detected in the urine of patients with active NCC.
CONCLUSIONS/SIGNIFICANCE: A newly developed recombinant monoclonal antibody-based Ts Ag detection immunoassay is extremely sensitive in the detection of extra-parenchymal NCC and can be used to monitor the success of treatment in the CSF, serum/plasma and urine. The ability to produce recombinant TsG10 at scale should enable use of this antigen detection immunoassay wherever NCC is endemic.
ClinicalTrials.gov Identifiers: NCT00001205 - & NCT00001645.
抗原检测可用于某些类型的神经囊尾蚴病（NCC）的诊断和疾病监测，但由于专有试剂和/或试剂盒的可用性，检测的可及性受到限制。
方法/主要发现：三个先前鉴定的分泌 IgM 的杂交瘤，其 IgM 产物显示出对猪带绦虫的特异性，经历了可变重链和轻链测序以及同种型转换为鼠 IgG。对这些重组表达的 IgG 抗 Ts 杂交瘤进行筛选，发现一个（TsG10）对粗制绦虫抗原具有最高亲和力。然后，TsG10 被用作夹心抗原检测免疫测定中的捕获抗体，与高滴度多克隆抗 Ts 抗体或生物素化 TsG10（称为 TsG10*bt）结合使用。使用来自活动性 NCC 患者和未感染 NCC 患者的血清、血浆和 CSF 样本，ROC 曲线分析表明，TsG10-TsG10-*bt 测定法在检测已知抗原阳性的样本时达到了 98%的灵敏度和 100%的特异性，优于基于多克隆抗体的测定法（灵敏度为 93%，特异性为 100%）。通过比较来自具有明确诊断的脑外 NCC 的特征良好的患者的配对 CSF（n=10）或血浆/血清（n=19）样本中 Ts 抗原（Ag）的水平，除了 2 个样本（1 个来自 CSF，1 个来自血浆）外，所有样本均无法检测到。新测定法检测到的 Ag 水平与商业测定法发现的水平高度相关（r=0.98）。初步研究表明，这种抗原可以在活动性 NCC 患者的尿液中检测到。
结论/意义：新开发的基于重组单克隆抗体的 Ts Ag 检测免疫测定法在检测脑外 NCC 方面非常敏感，可用于监测 CSF、血清/血浆和尿液中治疗的成功。能够大规模生产重组 TsG10 应该能够使这种抗原检测免疫测定法在 NCC 流行的任何地方都得到应用。
ClinicalTrials.gov 标识符：NCT00001205-和 NCT00001645。
