{"title":"SHAPU 的先进诊断方法：元基因组测序和白细胞介素分析","authors":"Ranju Kharel Sitaula , Rajeev Shrestha , Pratap Karki , Haramaya Gurung , Anadi Khatri KC , Nishan Katuwal , Navin Adhikari , Rajendra Bhatta , Eliya Shrestha , Indraman Maharjan , Pursottam Joshi , Biraj Man Karmacharya , Angira Shrestha , Ananda K Sharma , Sagun Narayan Joshi , Madan Prasad Upadhaya","doi":"10.1016/j.ajoint.2024.100056","DOIUrl":null,"url":null,"abstract":"<div><h3>Purpose</h3><p>Seasonal Hyperacute Panuveitis (SHAPU) is an ocular emergency where the genesis between infective and immunological is under investigation. This is the first report in the literature that attempts to explore the unidentified cause using next-generation metagenomic sequencing and immunomarker analysis.</p></div><div><h3>Design</h3><p>Cross-sectional quantitative study.</p></div><div><h3>Methods</h3><p>In this study, vitreous samples were collected from the SHAPU cases during the outbreak of 2019/2020. The vitreous samples of 10 random cases got selected for metagenomic next-generation sequencing. And 5 vitreous samples ran for immune analysis of levels of interleukin (IL) 6 and 10 using the ELISA technique. The ocular samples were extracted and sequenced on Illumina iSeq100 and analyzed through CZ ID platform. The true hits, for the suspected causative organisms were confirmed using qPCR through designed primers specific for that particular organism.</p></div><div><h3>Results</h3><p>A total of 30 eyes of 30 patients were included. Upon sequencing the DNA libraries, hits were observed for Streptococcus pneumoniae in 2/10 cases, confirmed by qPCR. IL-6 level was increased in 5/5 cases with IL-6/IL-10 greater than 1.</p></div><div><h3>Conclusion</h3><p>We report the presence of S. pneumoniae for the first time using a metagenomics analysis and increased IL-6 to IL-10 ratio via immune-analysis, indicating that SHAPU could have both infective and immunological facets on its causation.</p></div>","PeriodicalId":100071,"journal":{"name":"AJO International","volume":"1 3","pages":"Article 100056"},"PeriodicalIF":0.0000,"publicationDate":"2024-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S295025352400056X/pdfft?md5=f28712d9614b71c191f8a3591b4e603f&pid=1-s2.0-S295025352400056X-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Advanced diagnostic approaches for SHAPU: Metagenomic sequencing and interleukin analysis\",\"authors\":\"Ranju Kharel Sitaula , Rajeev Shrestha , Pratap Karki , Haramaya Gurung , Anadi Khatri KC , Nishan Katuwal , Navin Adhikari , Rajendra Bhatta , Eliya Shrestha , Indraman Maharjan , Pursottam Joshi , Biraj Man Karmacharya , Angira Shrestha , Ananda K Sharma , Sagun Narayan Joshi , Madan Prasad Upadhaya\",\"doi\":\"10.1016/j.ajoint.2024.100056\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Purpose</h3><p>Seasonal Hyperacute Panuveitis (SHAPU) is an ocular emergency where the genesis between infective and immunological is under investigation. This is the first report in the literature that attempts to explore the unidentified cause using next-generation metagenomic sequencing and immunomarker analysis.</p></div><div><h3>Design</h3><p>Cross-sectional quantitative study.</p></div><div><h3>Methods</h3><p>In this study, vitreous samples were collected from the SHAPU cases during the outbreak of 2019/2020. The vitreous samples of 10 random cases got selected for metagenomic next-generation sequencing. And 5 vitreous samples ran for immune analysis of levels of interleukin (IL) 6 and 10 using the ELISA technique. The ocular samples were extracted and sequenced on Illumina iSeq100 and analyzed through CZ ID platform. The true hits, for the suspected causative organisms were confirmed using qPCR through designed primers specific for that particular organism.</p></div><div><h3>Results</h3><p>A total of 30 eyes of 30 patients were included. Upon sequencing the DNA libraries, hits were observed for Streptococcus pneumoniae in 2/10 cases, confirmed by qPCR. IL-6 level was increased in 5/5 cases with IL-6/IL-10 greater than 1.</p></div><div><h3>Conclusion</h3><p>We report the presence of S. pneumoniae for the first time using a metagenomics analysis and increased IL-6 to IL-10 ratio via immune-analysis, indicating that SHAPU could have both infective and immunological facets on its causation.</p></div>\",\"PeriodicalId\":100071,\"journal\":{\"name\":\"AJO International\",\"volume\":\"1 3\",\"pages\":\"Article 100056\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-07-15\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S295025352400056X/pdfft?md5=f28712d9614b71c191f8a3591b4e603f&pid=1-s2.0-S295025352400056X-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"AJO International\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S295025352400056X\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"AJO International","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S295025352400056X","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
目的季节性高急性葡萄膜炎（SHAPU）是一种眼部急症，感染性和免疫性之间的成因正在研究之中。这是文献中第一份试图利用新一代元基因组测序和免疫标志物分析来探索未确定病因的报告。设计横断面定量研究。方法本研究收集了2019/2020年疫情爆发期间SHAPU病例的玻璃体样本。随机选取 10 例病例的玻璃体样本进行元基因组下一代测序。5份玻璃体样本采用ELISA技术对白细胞介素（IL）6和10的水平进行免疫分析。眼部样本在 Illumina iSeq100 上进行提取和测序，并通过 CZ ID 平台进行分析。通过为特定生物设计的特异引物，使用 qPCR 对疑似致病生物的真正命中进行确认。对 DNA 文库进行测序后，2/10 的病例发现了肺炎链球菌，并通过 qPCR 进行了确认。结论我们首次通过元基因组学分析报告了肺炎链球菌的存在，并通过免疫分析报告了IL-6与IL-10比值的升高，这表明SHAPU可能同时具有感染和免疫两方面的致病因素。
Advanced diagnostic approaches for SHAPU: Metagenomic sequencing and interleukin analysis
Purpose
Seasonal Hyperacute Panuveitis (SHAPU) is an ocular emergency where the genesis between infective and immunological is under investigation. This is the first report in the literature that attempts to explore the unidentified cause using next-generation metagenomic sequencing and immunomarker analysis.
Design
Cross-sectional quantitative study.
Methods
In this study, vitreous samples were collected from the SHAPU cases during the outbreak of 2019/2020. The vitreous samples of 10 random cases got selected for metagenomic next-generation sequencing. And 5 vitreous samples ran for immune analysis of levels of interleukin (IL) 6 and 10 using the ELISA technique. The ocular samples were extracted and sequenced on Illumina iSeq100 and analyzed through CZ ID platform. The true hits, for the suspected causative organisms were confirmed using qPCR through designed primers specific for that particular organism.
Results
A total of 30 eyes of 30 patients were included. Upon sequencing the DNA libraries, hits were observed for Streptococcus pneumoniae in 2/10 cases, confirmed by qPCR. IL-6 level was increased in 5/5 cases with IL-6/IL-10 greater than 1.
Conclusion
We report the presence of S. pneumoniae for the first time using a metagenomics analysis and increased IL-6 to IL-10 ratio via immune-analysis, indicating that SHAPU could have both infective and immunological facets on its causation.
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