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Simultaneous Detection of HIV-1 and TB Peptides with high sensitivity by Immuno-Affinity LC–MS/MS

Lin Li, PhD, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital


Li Jiang, PhD, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital; Director, Sichuan Clinical Laboratory Center


Tony Ye Hu, PhD, Tulane University Weatherhead Presidential Chair in Biotechnology Innovation; Director, Center for Cellular and Molecular Diagnostics; Fellow, National Academy of Inventors


Living with HIV significantly increases the risk of developing tuberculosis (TB), which remains a leading cause of illness and death among HIV-positive individuals. Current guidelines stress the importance of regular screening for TB during HIV care visits, as well as HIV screening for patients with TB. Integrating services for both diseases is crucial in mitigating their impact, emphasizing the need for innovative approaches in diagnosis and treatment monitoring.


In response to this pressing need, a team of researchers that are NACCCA members (Drs. Lin Li, Li Jiang and Tony Ye Hu) has developed and validated a novel assay of immuno-affinity liquid chromatography–tandem mass spectrometry (ILM) and published on Clinical Chemistry. This assay simultaneously quantifies specific peptides derived from HIV-1 p24 and Mycobacterium tuberculosis (Mtb) 10-kDa culture filtrate protein (CFP10) in serum samples.


The results of the study, which involved cohorts of both adults and children with and without HIV and TB, demonstrated the effectiveness of the ILM assay. It exhibited good precision and accuracy in detecting both p24 and CFP10 peptides, with high sensitivity and specificity for diagnosing HIV-1 and TB. Importantly, the assay detected HIV-1 infection earlier than existing commercial immunoassays, highlighting its potential for early detection.


Furthermore, longitudinal analysis of serum samples from children with HIV-1 and TB provided valuable insights into treatment response. The levels of p24 and CFP10 distinguished individuals who responded well to TB treatment from those who did not, as well as those who developed TB immune reconstitution inflammatory syndrome. This ability to monitor treatment response could significantly aid in the management of both diseases.


The findings of this study offer promising implications for the early diagnosis and treatment monitoring of HIV and TB. By providing a means to simultaneously detect both diseases and assess treatment response, the ILM assay has the potential to streamline healthcare delivery and improve patient outcomes. Integration of TB and HIV services is essential in addressing the dual burden of these diseases, and the development of innovative diagnostic tools like the ILM assay represents a significant step forward in achieving this goal.

 

利用免疫亲和液相色谱-串联质谱法高灵敏度地同时检测 HIV-1 和结核分枝杆菌肽


作者:


李霖博士,四川省医学科学院, 四川省人民医院


蒋黎教授,博士,四川省医学科学院, 四川省人民医院;四川省临床检验中心主任


胡晔教授,博士,杜兰大学生物技术创新校长讲席教授;细胞与分子诊断中心主任;美国国家发明家学院院士


艾滋病病毒感染者罹患结核(TB)的风险大大增加,而结核仍然是艾滋病病毒抗体阳性者患病和死亡的主要原因。目前的指导方针强调了在艾滋病毒患者就诊期间定期筛查肺结核以及对结核患者进行艾滋病毒筛查的重要性。整合针对这两种疾病的筛查对于减轻其影响至关重要,这强调了在诊断和治疗监测方面采用创新方法的必要性。


为了满足这一迫切需求,一个由 NACCCA 会员组成的研究团队(李琳博士、蒋黎 博士和胡晔博士)开发并验证了一种新型的免疫亲和液相色谱-串联质谱(ILM)检测方法并且发表在临床化学期刊。该检测方法可同时定量检测血清样本中来自 HIV-1 p24 和结核分枝杆菌(Mtb)10-kDa 培养滤液蛋白(CFP10)的特异性多肽。


这项研究的对象包括感染或未感染艾滋病毒和结核病的成人和儿童,研究结果证明了 ILM 检测法的有效性。它在检测 p24 和 CFP10 肽方面表现出良好的精确性和准确性,在诊断 HIV-1 和结核方面具有很高的灵敏度和特异性。重要的是,该检测方法比现有的商业免疫测定方法更早地检测出 HIV-1 感染,突出了其早期检测的潜力。


此外,对感染 HIV-1 和结核病的儿童血清样本进行的纵向分析为了解治疗反应提供了宝贵的信息。p24 和 CFP10 的水平可以区分对结核治疗反应良好的患者和反应不佳的患者,以及出现结核免疫重建炎症综合征的患者。这种监测治疗反应的能力将大大有助于这两种疾病的治疗。


这项研究的结果为艾滋病和结核病的早期诊断和治疗监测带来了希望。通过提供一种同时检测两种疾病和评估治疗反应的方法,ILM 检测法有可能简化医疗服务并改善患者的治疗效果。整合结核病和艾滋病服务对于解决这些疾病带来的双重负担至关重要,而像 ILM 检测法这样的创新诊断工具的开发则是实现这一目标的重要一步。

 

References

Lin Li, Christopher J Lyon, Sylvia M LaCourse, Wenshu Zheng, Joshua Stern, Jaclyn N Escudero, Wilfred Bundi Murithi, Lilian Njagi, Grace John-Stewart, Thomas R Hawn, Videlis Nduba, Wael Abdelgaliel, Thomas Tombler, David Horne, Li Jiang, Tony Y Hu.

Sensitive Blood-Based Detection of HIV-1 and Mycobacterium tuberculosis Peptides for Disease Diagnosis by Immuno-Affinity Liquid Chromatography–Tandem Mass Spectrometry: A Method Development and Proof-of-Concept Study.

Clinical Chemistry, Volume 69, Issue 12, December 2023, Pages 1409–1419.

 

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