Chaturvedi Chandra P, Hashim Zia, Tripathy Naresh K
Department of Hematology, Stem Cell Research Center, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, IND.
Department of Pulmonary Medicine, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, IND.
Cureus. 2022 Jun 17;14(6):e26025. doi: 10.7759/cureus.26025. eCollection 2022 Jun.
Background and objectives The mannoprotein lateral flow assay (MP-LFA) or Aspergillus-specific lateral flow device (AspLFD) is a novel rapid test for point-of-care diagnosis (PoC) of invasive aspergillosis (IA), but its routine clinical application is hampered due to low sensitivity (S) of the assay in serum. Therefore, this study aimed to develop a new method to enhance the S of the serum MP-LFA. Methodology In the new method (Tripathy method), we used direct heating of the serum without any dilution at 120C for 15 minutes to purify the mannoprotein (MP) antigen of the Aspergillus. The MP-enriched serum supernatant obtained after centrifugation was loaded in an LFD cassette, and the results were read after 20 minutes using a digital cube reader. In parallel to our new method, AspLFD was performed according to the manufacturer's instructions. The diagnostic performance of the two methods was evaluated using paired sera of true IA patients (IA, n=18) and healthy subjects (controls, n=20). The positivity of the two methods was also evaluated in the sera of leukemia patients with possible/probable IA (possible/probable IA; n=23). Results The Tripathy method had a significantly higher sensitivity (88.9% versus 55.5%; p<0.05) and diagnostic odds ratio (72.0 versus 23.7) than the standard AspLFD method. In receiver operating characteristic curve analysis for differentiation between IA patients and controls, although the Tripathy method (area under curve; AUC: 0.894, p<0.001) and AspLFD method (AUC: 0.753, p<0.001) were significantly associated with IA, the AUC of the Tripathy method was significantly higher than that of the AspLFD method (0.894 versus 0.753; p<0.05). In the sera of possible/probable IA, MP-LFA by the Tripathy method had a significantly higher rate of positivity than the AspLFD method (39.0% versus 21.7%; p<0.05). Conclusion Our data show that the Tripathy method is a highly sensitive method of MP-LFA for the PoC diagnosis of IA in clinical settings.
甘露糖蛋白侧向流动分析法（MP-LFA）或曲霉特异性侧向流动检测装置（AspLFD）是一种用于侵袭性曲霉病（IA）即时诊断（PoC）的新型快速检测方法，但由于该检测方法在血清中的敏感性（S）较低，其常规临床应用受到阻碍。因此，本研究旨在开发一种新方法来提高血清MP-LFA的敏感性。方法：在新方法（特里帕蒂方法）中，我们在120℃下直接加热血清15分钟，且不进行任何稀释，以纯化曲霉的甘露糖蛋白（MP）抗原。将离心后获得的富含MP的血清上清液加载到LFD检测卡中，并使用数字立方读数器在20分钟后读取结果。与我们的新方法并行，按照制造商的说明进行AspLFD检测。使用确诊IA患者（IA，n = 18）和健康受试者（对照组，n = 20）的配对血清评估这两种方法的诊断性能。还在可能/疑似IA的白血病患者血清中评估了这两种方法的阳性率（可能/疑似IA；n = 23）。结果：与标准的AspLFD方法相比，特里帕蒂方法具有显著更高的敏感性（88.9%对55.5%；p<0.05）和诊断比值比（72.0对23.7）。在用于区分IA患者和对照组的受试者工作特征曲线分析中，尽管特里帕蒂方法（曲线下面积；AUC：0.894，p<0.001）和AspLFD方法（AUC：0.753，p<0.001）均与IA显著相关，但特里帕蒂方法的AUC显著高于AspLFD方法（0.894对0.753；p<0.05）。在可能/疑似IA的血清中，采用特里帕蒂方法的MP-LFA阳性率显著高于AspLFD方法（39.0%对21.7%；p<0.05）。结论：我们的数据表明，特里帕蒂方法是一种用于临床环境中IA即时诊断的高度敏感的MP-LFA方法。
