对从丙型肝炎患者血液中提取的RNA样本进行表面增强拉曼光谱分析以定量病毒载量。
Surface enhanced Raman spectroscopy of RNA samples extracted from blood of hepatitis C patients for quantification of viral loads.
作者信息
Nasir Saira, Majeed Muhammad Irfan, Nawaz Haq, Rashid Nosheen, Ali Saqib, Farooq Sidra, Kashif Muhammad, Rafiq Sidra, Bano Saira, Ashraf Muhammad Naeem, Abubakar Muhammad, Ahmad Shamsheer, Rehman Asma, Amin Imran
机构信息
Department of Chemistry, University of Agriculture Faisalabad, 38040, Pakistan.
Department of Chemistry, University of Agriculture Faisalabad, 38040, Pakistan.
出版信息
Photodiagnosis Photodyn Ther. 2021 Mar;33:102152. doi: 10.1016/j.pdpdt.2020.102152. Epub 2020 Dec 30.
BACKGROUND
Raman spectroscopy is a promising technique to analyze the body fluids for the purpose of non-invasive disease diagnosis.
OBJECTIVES
To develop a surface-enhanced Raman spectroscopy (SERS) based method for qualitative and quantitative analysis of HCV from blood samples.
METHODS
SERS was employed to characterize the Hepatitis C viral RNA extracted from different blood samples of hepatitis C virus (HCV) infected patients with predetermined viral loads in comparison with total RNA of healthy individuals. The SERS measurements were performed on 27 extracted RNA samples including low viral loads, medium viral loads, high viral loads and healthy/negative viral load samples. For this purpose, silver nanoparticles (Ag NPs) were used as SERS substrates. Furthermore, multivariate data analysis technique, Principal Component Analysis (PCA) and Partial Least Square Regression (PLSR) were also performed on SERS spectral data.
RESULTS
The SERS spectral features due to biochemical changes in the extracted RNA samples associated with the increasing viral loads were established which could be employed for HCV diagnostic purpose. PCA was found helpful for the differentiation between Raman spectral data of RNA extracted from hepatitis infected and healthy blood samples. PLSR model is established for the determination of viral loads in HCV positive RNA samples with 99 % accuracy.
CONCLUSION
SERS can be employed for qualitative and quantitative analysis of HCV from blood samples.
背景
拉曼光谱法是一种很有前景的技术，可用于分析体液以实现非侵入性疾病诊断。
目的
开发一种基于表面增强拉曼光谱（SERS）的方法，用于对血液样本中的丙型肝炎病毒（HCV）进行定性和定量分析。
方法
采用SERS对从丙型肝炎病毒（HCV）感染患者的不同血液样本中提取的丙型肝炎病毒RNA进行表征，这些患者具有预先确定的病毒载量，并与健康个体的总RNA进行比较。对27个提取的RNA样本进行了SERS测量，包括低病毒载量、中等病毒载量、高病毒载量以及健康/阴性病毒载量样本。为此，使用银纳米颗粒（Ag NPs）作为SERS底物。此外，还对SERS光谱数据进行了多元数据分析技术，即主成分分析（PCA）和偏最小二乘回归（PLSR）。
结果
确定了与病毒载量增加相关的提取RNA样本中生化变化引起的SERS光谱特征，可用于HCV诊断。发现PCA有助于区分从肝炎感染血液样本和健康血液样本中提取的RNA的拉曼光谱数据。建立了PLSR模型，用于测定HCV阳性RNA样本中的病毒载量，准确率达99%。
结论
SERS可用于对血液样本中的HCV进行定性和定量分析。
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