Dilek Çam Derin, Enes Gültekin, Elif Gündüz, Barış Otlu
{"title":"通过横向流动检测法快速检测 SARS-CoV-2 的 6 对适配体-适配体的比较","authors":"Dilek Çam Derin, Enes Gültekin, Elif Gündüz, Barış Otlu","doi":"10.1093/jaoacint/qsae004","DOIUrl":null,"url":null,"abstract":"Background SARS-CoV-2 is a threat for humanity. Both the Spike (S) protein and its receptor binding domain (sRBD) are extensively used for the rapid detection. Although real time reverse transcription polymerase chain reaction (rRT-PCR) is mostly used method for the molecular detection of SARS-CoV-2, rapid assays for antigenic detection is always needed. Lateral flow assays (LFAs) are the most commonly used tests for this purpose and aptamers having stability and long shelf life are used as capture reagents. Objective This study aimed to develop the LFAs based on the aptamer pairs for the antigenic detection of SARS-CoV-2 with naked eye. Methods Gold nanoparticles (AuNPs) were used as label and six sandwich models by three different aptamers were prepared using 4 μM and 8 μM probes and two kinds of membranes for developing the LFAs. Results 8 μM probe concentration and M2 membrane showed the best recognition of both the synthetic sRBD and SARS-CoV-2 coming from the naso/oropharingeal swabs by designed LFAs as 100% sensitivity and 93,3% specifity compared to the antibody detecting LFAs. Conclusions Our developed strip assays based on aptamer pairs recognized the target, directly in a 5-6 minutes with naked eye. It was also concluded that aptamer pairs, membrane types, assay buffers and probe concentrations have significant role in the detection of SARS-CoV-2 by LFAs. Highlights The detection of SARS-CoV-2 in clinical samples was demonstrated with the best aptamer pairs, sensitively and selectively among the designed 6 aptamer pairs for LFAs. Developed LFAs can be an alternative method to the conventional antibody based LFAs for SARS-CoV-2 detection.","PeriodicalId":15003,"journal":{"name":"Journal of AOAC International","volume":null,"pages":null},"PeriodicalIF":1.7000,"publicationDate":"2024-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Comparison of 6 aptamer-aptamer pairs on rapid detection of SARS-CoV-2 by lateral flow assay\",\"authors\":\"Dilek Çam Derin, Enes Gültekin, Elif Gündüz, Barış Otlu\",\"doi\":\"10.1093/jaoacint/qsae004\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Background SARS-CoV-2 is a threat for humanity. Both the Spike (S) protein and its receptor binding domain (sRBD) are extensively used for the rapid detection. Although real time reverse transcription polymerase chain reaction (rRT-PCR) is mostly used method for the molecular detection of SARS-CoV-2, rapid assays for antigenic detection is always needed. Lateral flow assays (LFAs) are the most commonly used tests for this purpose and aptamers having stability and long shelf life are used as capture reagents. Objective This study aimed to develop the LFAs based on the aptamer pairs for the antigenic detection of SARS-CoV-2 with naked eye. Methods Gold nanoparticles (AuNPs) were used as label and six sandwich models by three different aptamers were prepared using 4 μM and 8 μM probes and two kinds of membranes for developing the LFAs. Results 8 μM probe concentration and M2 membrane showed the best recognition of both the synthetic sRBD and SARS-CoV-2 coming from the naso/oropharingeal swabs by designed LFAs as 100% sensitivity and 93,3% specifity compared to the antibody detecting LFAs. Conclusions Our developed strip assays based on aptamer pairs recognized the target, directly in a 5-6 minutes with naked eye. It was also concluded that aptamer pairs, membrane types, assay buffers and probe concentrations have significant role in the detection of SARS-CoV-2 by LFAs. Highlights The detection of SARS-CoV-2 in clinical samples was demonstrated with the best aptamer pairs, sensitively and selectively among the designed 6 aptamer pairs for LFAs. Developed LFAs can be an alternative method to the conventional antibody based LFAs for SARS-CoV-2 detection.\",\"PeriodicalId\":15003,\"journal\":{\"name\":\"Journal of AOAC International\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.7000,\"publicationDate\":\"2024-01-11\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of AOAC International\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.1093/jaoacint/qsae004\",\"RegionNum\":4,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"CHEMISTRY, ANALYTICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of AOAC International","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1093/jaoacint/qsae004","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
Comparison of 6 aptamer-aptamer pairs on rapid detection of SARS-CoV-2 by lateral flow assay
Background SARS-CoV-2 is a threat for humanity. Both the Spike (S) protein and its receptor binding domain (sRBD) are extensively used for the rapid detection. Although real time reverse transcription polymerase chain reaction (rRT-PCR) is mostly used method for the molecular detection of SARS-CoV-2, rapid assays for antigenic detection is always needed. Lateral flow assays (LFAs) are the most commonly used tests for this purpose and aptamers having stability and long shelf life are used as capture reagents. Objective This study aimed to develop the LFAs based on the aptamer pairs for the antigenic detection of SARS-CoV-2 with naked eye. Methods Gold nanoparticles (AuNPs) were used as label and six sandwich models by three different aptamers were prepared using 4 μM and 8 μM probes and two kinds of membranes for developing the LFAs. Results 8 μM probe concentration and M2 membrane showed the best recognition of both the synthetic sRBD and SARS-CoV-2 coming from the naso/oropharingeal swabs by designed LFAs as 100% sensitivity and 93,3% specifity compared to the antibody detecting LFAs. Conclusions Our developed strip assays based on aptamer pairs recognized the target, directly in a 5-6 minutes with naked eye. It was also concluded that aptamer pairs, membrane types, assay buffers and probe concentrations have significant role in the detection of SARS-CoV-2 by LFAs. Highlights The detection of SARS-CoV-2 in clinical samples was demonstrated with the best aptamer pairs, sensitively and selectively among the designed 6 aptamer pairs for LFAs. Developed LFAs can be an alternative method to the conventional antibody based LFAs for SARS-CoV-2 detection.
期刊介绍:
The Journal of AOAC INTERNATIONAL publishes the latest in basic and applied research in analytical sciences related to foods, drugs, agriculture, the environment, and more. The Journal is the method researchers'' forum for exchanging information and keeping informed of new technology and techniques pertinent to regulatory agencies and regulated industries.