{"id":4383,"date":"2025-12-17T08:48:15","date_gmt":"2025-12-17T01:48:15","guid":{"rendered":"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/?p=4383"},"modified":"2025-12-17T08:49:23","modified_gmt":"2025-12-17T01:49:23","slug":"production-of-recombinant-electron-transfer-flavoprotein-beta-subunit-protein-and-its-application-in-a-lateral-flow-assay-for-early-diagnosis-of-leptospirosis","status":"publish","type":"post","link":"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/production-of-recombinant-electron-transfer-flavoprotein-beta-subunit-protein-and-its-application-in-a-lateral-flow-assay-for-early-diagnosis-of-leptospirosis\/","title":{"rendered":"Production of recombinant electron transfer flavoprotein beta subunit protein and its application in a lateral flow assay for early diagnosis of leptospirosis"},"content":{"rendered":"<h2 class=\"title\">Abstract<\/h2>\n<div id=\"eng-abstract\" class=\"abstract-content selected\">\n<p>Leptospirosis is a major cause of acute febrile illness, often presenting with non-specific symptoms that can lead to misdiagnosis. Early laboratory diagnosis is essential for confirmation to avoid misdiagnosis and ensure appropriate management. This study aimed to identify and produce a recombinant protein, approximately 25 kDa, with high antigenicity for diagnostic applications. The 25 kDa protein from Leptospira interrogans was identified as electron transfer flavoprotein beta subunits (Etf\u03b2) and exhibited 98% nucleotide and 99% amino acid homology to the reference strain. Lateral flow assays (LFAs) using recombinant Etf\u03b2 (rEtf\u03b2) as antigens were developed to detect specific antibodies, namely rEtf\u03b2-IgM and rEtf\u03b2-IgG, and evaluated their performance against the standard microscopic agglutination test (MAT). Testing 33 paired serum samples from confirmed leptospirosis cases and 24 controls revealed sensitivities of 69.7% for IgM and 57.6% for IgG. However, the combined assays yielded enhanced diagnostic accuracy, achieving a sensitivity of 94.0%, specificity of 95.8%, positive predictive value of 96.9%, negative predictive value of 92.0%, and percent agreement of 94.7% (kappa value of 0.89). Also, the combined LFAs demonstrated 66.7% in initial serum samples whose MAT results were negative, enhancing the capacity for early diagnosis. In conclusion, the developed rapid tests demonstrated strong diagnostic capability, particularly in early-phase leptospirosis, distinguishing between initial and recurrent infections. Importantly, rEtf\u03b2-IgG identified a subset of patients lacking detectable IgM. Thus, integrating rEtf\u03b2-IgM and rEtf\u03b2-IgG is recommended to improve sensitivity and accuracy in endemic populations. The rEtf\u03b2 is a promising target for future antigen-based diagnostic strategies for leptospirosis. The rEtf\u03b2 antigen shows promise as a target for future development of antigen-based diagnostic strategies for leptospirosis.<\/p>\n<p><a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/39969576\/\">https:\/\/pubmed.ncbi.nlm.nih.gov\/39969576\/<\/a><\/p>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Abstract Leptospirosis is a major cause of acute febrile illness, often presenting with non-specific symptoms that can lead to misdiagnosis. Early laboratory diagnosis is essential for confirmation to avoid misdiagnosis and ensure appropriate management. This study aimed to identify and produce a recombinant protein, approximately 25 kDa, with high antigenicity for diagnostic applications. The 25 [&#8230;]\n","protected":false},"author":6,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[18],"tags":[],"class_list":["post-4383","post","type-post","status-publish","format-standard","hentry","category-research-innovation-en"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.1.1 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Production of recombinant electron transfer flavoprotein beta subunit protein and its application in a lateral flow assay for early diagnosis of leptospirosis - \u0e20\u0e32\u0e04\u0e27\u0e34\u0e0a\u0e32\u0e0a\u0e35\u0e27\u0e40\u0e04\u0e21\u0e35 \u0e04\u0e13\u0e30\u0e41\u0e1e\u0e17\u0e22\u0e28\u0e32\u0e2a\u0e15\u0e23\u0e4c\u0e28\u0e34\u0e23\u0e34\u0e23\u0e32\u0e0a\u0e1e\u0e22\u0e32\u0e1a\u0e32\u0e25 \u0e21\u0e2b\u0e32\u0e27\u0e34\u0e17\u0e22\u0e32\u0e25\u0e31\u0e22\u0e21\u0e2b\u0e34\u0e14\u0e25<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/production-of-recombinant-electron-transfer-flavoprotein-beta-subunit-protein-and-its-application-in-a-lateral-flow-assay-for-early-diagnosis-of-leptospirosis\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Production of recombinant electron transfer flavoprotein beta subunit protein and its application in a lateral flow assay for early diagnosis of leptospirosis - \u0e20\u0e32\u0e04\u0e27\u0e34\u0e0a\u0e32\u0e0a\u0e35\u0e27\u0e40\u0e04\u0e21\u0e35 \u0e04\u0e13\u0e30\u0e41\u0e1e\u0e17\u0e22\u0e28\u0e32\u0e2a\u0e15\u0e23\u0e4c\u0e28\u0e34\u0e23\u0e34\u0e23\u0e32\u0e0a\u0e1e\u0e22\u0e32\u0e1a\u0e32\u0e25 \u0e21\u0e2b\u0e32\u0e27\u0e34\u0e17\u0e22\u0e32\u0e25\u0e31\u0e22\u0e21\u0e2b\u0e34\u0e14\u0e25\" \/>\n<meta property=\"og:description\" content=\"Abstract Leptospirosis is a major cause of acute febrile illness, often presenting with non-specific symptoms that can lead to misdiagnosis. Early laboratory diagnosis is essential for confirmation to avoid misdiagnosis and ensure appropriate management. This study aimed to identify and produce a recombinant protein, approximately 25 kDa, with high antigenicity for diagnostic applications. The 25 [...]\" \/>\n<meta property=\"og:url\" content=\"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/production-of-recombinant-electron-transfer-flavoprotein-beta-subunit-protein-and-its-application-in-a-lateral-flow-assay-for-early-diagnosis-of-leptospirosis\/\" \/>\n<meta property=\"og:site_name\" content=\"\u0e20\u0e32\u0e04\u0e27\u0e34\u0e0a\u0e32\u0e0a\u0e35\u0e27\u0e40\u0e04\u0e21\u0e35 \u0e04\u0e13\u0e30\u0e41\u0e1e\u0e17\u0e22\u0e28\u0e32\u0e2a\u0e15\u0e23\u0e4c\u0e28\u0e34\u0e23\u0e34\u0e23\u0e32\u0e0a\u0e1e\u0e22\u0e32\u0e1a\u0e32\u0e25 \u0e21\u0e2b\u0e32\u0e27\u0e34\u0e17\u0e22\u0e32\u0e25\u0e31\u0e22\u0e21\u0e2b\u0e34\u0e14\u0e25\" \/>\n<meta property=\"article:publisher\" content=\"https:\/\/www.facebook.com\/SIBBprogram\/\" \/>\n<meta property=\"article:published_time\" content=\"2025-12-17T01:48:15+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2025-12-17T01:49:23+00:00\" \/>\n<meta name=\"author\" content=\"Poorintra Sakuna\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Written by\" \/>\n\t<meta name=\"twitter:data1\" content=\"Poorintra Sakuna\" \/>\n\t<meta name=\"twitter:label2\" content=\"Est. reading time\" \/>\n\t<meta name=\"twitter:data2\" content=\"2 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\/\/schema.org\",\"@graph\":[{\"@type\":\"Article\",\"@id\":\"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/production-of-recombinant-electron-transfer-flavoprotein-beta-subunit-protein-and-its-application-in-a-lateral-flow-assay-for-early-diagnosis-of-leptospirosis\/#article\",\"isPartOf\":{\"@id\":\"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/production-of-recombinant-electron-transfer-flavoprotein-beta-subunit-protein-and-its-application-in-a-lateral-flow-assay-for-early-diagnosis-of-leptospirosis\/\"},\"author\":{\"name\":\"Poorintra Sakuna\",\"@id\":\"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/#\/schema\/person\/75170c4a414d080cfd0b7428b2e0d11a\"},\"headline\":\"Production of recombinant electron transfer flavoprotein beta subunit protein and its application in a lateral flow assay for early diagnosis of leptospirosis\",\"datePublished\":\"2025-12-17T01:48:15+00:00\",\"dateModified\":\"2025-12-17T01:49:23+00:00\",\"mainEntityOfPage\":{\"@id\":\"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/production-of-recombinant-electron-transfer-flavoprotein-beta-subunit-protein-and-its-application-in-a-lateral-flow-assay-for-early-diagnosis-of-leptospirosis\/\"},\"wordCount\":284,\"publisher\":{\"@id\":\"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/#organization\"},\"articleSection\":[\"Research Innovation\"],\"inLanguage\":\"en-US\"},{\"@type\":\"WebPage\",\"@id\":\"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/production-of-recombinant-electron-transfer-flavoprotein-beta-subunit-protein-and-its-application-in-a-lateral-flow-assay-for-early-diagnosis-of-leptospirosis\/\",\"url\":\"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/production-of-recombinant-electron-transfer-flavoprotein-beta-subunit-protein-and-its-application-in-a-lateral-flow-assay-for-early-diagnosis-of-leptospirosis\/\",\"name\":\"Production of recombinant electron transfer flavoprotein beta subunit protein and its application in a lateral flow assay for early diagnosis of leptospirosis - 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