Laboratory of Molecular Virology, Oswaldo Cruz Institute/Fiocruz, Rio de Janeiro, Brazil.
Hematology Department, Gaffrée & Guinle Universitary Hospital, Rio de Janeiro State Federal University/UniRio, Rio de Janeiro, Brazil.
J Appl Microbiol. 2018 Oct;125(4):1208-1215. doi: 10.1111/jam.14024. Epub 2018 Jul 30.
To improve RT-qPCR with an internal control and a synthetic standard curve to detect HEV in HIV co-infected patients.
A single-stranded RNA (ssRNA) and a double-stranded DNA (dsDNA) synthetic curve were designed, compared to the international reference panel for HEV genotypes, and tested to quantify and detect a reference panel for HEV genotypes. The detection limit of the RNA synthetic curve (50 copies per ml) was better than the DNA synthetic curve (100 copies per ml) and the WHO standard curve (250 copies per ml). Then, 280 serum samples from HIV-positive patients were tested for HEV RNA, which was detected in 3·6% of serum samples. The viral load ranged from 2 × 10 copies per ml to 4·78 × 10 copies per ml. HEV IgM/IgG antibodies were not detected in the RNA-positive patients. Sequencing analysis of HEV showed that the virus belongs to genotype 3 (HEV GT3).
Real-time PCR was a useful tool to estimate co-infection with HEV/HIV, even in patients with low viral loads and undetectable anti-HEV IgG and IgM antibodies.
Hepatitis E virus genotype 3 (HEV GT3) has been associated with silent chronic hepatitis and cirrhosis in HIV-positive subjects worldwide, but there is a lack of data on this co-infection in Brazil.
利用内参和合成标准曲线改进 RT-qPCR，以检测 HIV 合并感染患者中的 HEV。
设计了一条单链 RNA（ssRNA）和一条双链 DNA（dsDNA）合成曲线，与国际 HEV 基因型参考面板进行比较，并对其进行了测试，以定量和检测 HEV 基因型参考面板。RNA 合成曲线的检测限（50 拷贝/ml）优于 DNA 合成曲线（100 拷贝/ml）和世界卫生组织标准曲线（250 拷贝/ml）。然后，对 280 份来自 HIV 阳性患者的血清样本进行 HEV RNA 检测，其中 3.6%的血清样本检测到 HEV RNA。病毒载量范围为 2×10 拷贝/ml 至 4.78×10 拷贝/ml。在 RNA 阳性患者中未检测到 HEV IgM/IgG 抗体。HEV 的测序分析显示，该病毒属于基因型 3（HEV GT3）。
实时 PCR 是一种有用的工具，可以估计 HEV/HIV 合并感染，即使在病毒载量低且无法检测到抗 HEV IgG 和 IgM 抗体的患者中也是如此。
全球范围内，HEV 基因型 3（HEV GT3）已与 HIV 阳性患者中的无症状慢性肝炎和肝硬化相关，但在巴西，关于这种合并感染的数据很少。
