BSL-3 Laboratory, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, China.
Department of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, China.
Microbiol Spectr. 2022 Apr 27;10(2):e0149621. doi: 10.1128/spectrum.01496-21. Epub 2022 Mar 21.
In recent years, the chikungunya virus (CHIKV) has continued to spread from local epidemics to nonnative habitats until eventually reaching pandemic status. Nonendemic areas such as China have also emerged as potential epidemic areas of CHIKV. Serological detection of CHIKV is the key to diagnosing and controlling the prevalence of this virus. In this study, we review the progress of the serological detection of the envelope (E) protein in CHIKV, and we provide a novel research assay and ideas for the serological detection of CHIKV. The luciferase immunosorbent assay (LISA) does not require species-specific labeled secondary antibodies for detection, making it universally suitable for tracking samples from various animals or carriers. At present, most research on CHIKV antigen detection technology tends to combine two or more proteins to avoid the decrease in detection ability caused by antigen mutation. Our results indicate that two or more kinds of CHIKV E antigens combined with LISA detection can improve the detection rate of anti-CHIKV immunoglobulin G (IgG) antibodies in CHIKV-infected patient sera and detect antibodies in the early stage of infection accurately and sensitively. After 235 days of infection, the anti-CHIKV IgG antibodies could still be detected in CHIKV-infected patients. All serum samples were tested with a detection rate of 100% after combining various recombinant CHIKV E antigens. Our proposed CHIKV-specific LISA could be a useful tool for serum diagnosis of CHIKV infection and serum epidemic research in areas where CHIKV is endemic, which would help to manage potential epidemics in the future. At present, chikungunya virus (CHIKV) is still circulating in some parts of the world, and mutated strains have emerged, making it easier for the virus to spread among humans. With the continuous variation of CHIKV, its antigen variation leads to the decline of detection ability. In addition, the risk of transmission of CHIKV in areas where CHIKV is not endemic, such as China, has increased dramatically, which compels us to enhance the detection capacity of CHIKV and continuously monitor CHIKV antibody levels in the population. Real-time quantitative PCR (RT-PCR) detection technology will not be reliable when the infection time is chronic or in subclinical infection due to decreases in virus concentration, and an antibody detection technology must be adopted. In this study, multiple CHIKV envelope (E) antigens were used to detect anti-CHIKV IgG antibodies in serum for the first time. The new assay is characterized by convenient operation, high detection rate, and high sensitivity and has significance for early warning and monitoring. Moreover, it contributes to the prevention and control of CHIKV.
近年来，基孔肯雅病毒（CHIKV）不断从局部流行传播到非本地栖息地，最终达到大流行状态。中国等非流行地区也成为基孔肯雅病毒的潜在流行区。基孔肯雅病毒的血清学检测是诊断和控制该病毒流行的关键。本研究综述了基孔肯雅病毒包膜（E）蛋白血清学检测的进展，并为基孔肯雅病毒的血清学检测提供了一种新的研究方法和思路。荧光素酶免疫吸附试验（LISA）不需要物种特异性标记的二级抗体进行检测，因此普遍适用于跟踪来自各种动物或载体的样本。目前，大多数基孔肯雅病毒抗原检测技术的研究倾向于结合两种或两种以上的蛋白，以避免因抗原突变导致检测能力下降。我们的结果表明，两种或两种以上的基孔肯雅病毒 E 抗原与 LISA 检测相结合，可以提高基孔肯雅病毒感染患者血清中抗基孔肯雅病毒免疫球蛋白 G（IgG）抗体的检测率，并准确、敏感地检测到感染早期的抗体。感染后 235 天，仍可在基孔肯雅病毒感染患者中检测到抗基孔肯雅病毒 IgG 抗体。结合各种重组基孔肯雅病毒 E 抗原后，所有血清样本的检测率均为 100%。我们提出的基孔肯雅病毒特异性 LISA 可作为基孔肯雅病毒感染血清诊断和基孔肯雅病毒流行地区血清研究的有用工具，有助于未来管理潜在的流行。目前，基孔肯雅病毒（CHIKV）仍在世界部分地区传播，且出现了突变株，使病毒更容易在人群中传播。随着基孔肯雅病毒的不断变异，其抗原变异导致检测能力下降。此外，基孔肯雅病毒在非流行地区（如中国）的传播风险急剧增加，这迫使我们增强基孔肯雅病毒的检测能力，并持续监测人群中的基孔肯雅病毒抗体水平。实时定量 PCR（RT-PCR）检测技术在病毒浓度降低导致慢性感染或亚临床感染时不可靠，必须采用抗体检测技术。本研究首次使用多种基孔肯雅病毒包膜（E）抗原检测血清中抗基孔肯雅病毒 IgG 抗体。新的检测方法具有操作方便、检测率高、灵敏度高等特点，对早期预警和监测具有重要意义，有助于基孔肯雅病毒的预防和控制。
