{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"medRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T12:06:55Z","timestamp":1768565215691,"version":"3.49.0"},"posted":{"date-parts":[[2023,5,26]]},"group-title":"Infectious Diseases (except HIV\/AIDS)","reference-count":49,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,5,26]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec id=\"s21\">\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Neisseria gonorrhoeae<\/jats:italic>\n                    is one of the most common bacterial sexually transmitted infections. The emergence of antimicrobial-resistant\n                    <jats:italic>N. gonorrhoeae<\/jats:italic>\n                    is an urgent public health threat. Currently, diagnosis of\n                    <jats:italic>N. gonorrhoeae<\/jats:italic>\n                    infection requires expensive laboratory infrastructure, while antimicrobial susceptibility determination requires bacterial culture, both of which are infeasible in low-resource areas where prevalence is highest. Recent advances in molecular diagnostics, such as Specific High-sensitivity Enzymatic Reporter unLOCKing (SHERLOCK) using CRISPR-Cas13a and isothermal amplification, have the potential to provide low-cost detection of pathogen and antimicrobial resistance.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec id=\"s22\">\n                  <jats:title>Methods and Results<\/jats:title>\n                  <jats:p>\n                    We designed and optimized RNA guides and primer-sets for SHERLOCK assays capable of detecting\n                    <jats:italic>N. gonorrhoeae<\/jats:italic>\n                    via the\n                    <jats:italic>por<\/jats:italic>\n                    A gene and of predicting ciprofloxacin susceptibility via a single mutation in the gyrase A (\n                    <jats:italic>gyr<\/jats:italic>\n                    A) gene. We evaluated their performance using both synthetic DNA and purified\n                    <jats:italic>N. gonorrhoeae<\/jats:italic>\n                    isolates. For\n                    <jats:italic>por<\/jats:italic>\n                    A, we created both a fluorescence-based assay and lateral flow assay using a biotinylated FAM reporter. Both methods demonstrated sensitive detection of 14\n                    <jats:italic>N. gonorrhoeae<\/jats:italic>\n                    isolates and no cross-reactivity with 3 non-gonococcal\n                    <jats:italic>Neisseria<\/jats:italic>\n                    isolates. For\n                    <jats:italic>gyr<\/jats:italic>\n                    A, we created a fluorescence-based assay that correctly distinguished between 20 purified\n                    <jats:italic>N. gonorrhoeae<\/jats:italic>\n                    isolates with phenotypic ciprofloxacin resistance and 3 with phenotypic susceptibility. We confirmed the\n                    <jats:italic>gyr<\/jats:italic>\n                    A genotype predictions from the fluorescence-based assay with DNA sequencing, which showed 100% concordance for the isolates studied.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec id=\"s23\">\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>\n                    We report the development of Cas13a-based SHERLOCK assays that detect\n                    <jats:italic>N. gonorrhoeae<\/jats:italic>\n                    and differentiate ciprofloxacin-resistant isolates from ciprofloxacin-susceptible isolates.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2023.05.21.23290304","type":"posted-content","created":{"date-parts":[[2023,5,26]],"date-time":"2023-05-26T17:06:01Z","timestamp":1685120761000},"source":"Crossref","is-referenced-by-count":1,"title":["Development of Cas13a-based Assays for\n                  <i>Neisseria gonorrhoeae<\/i>\n                  Detection and Gyrase A Determination"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4077-3476","authenticated-orcid":false,"given":"Lao-Tzu","family":"Allan-Blitz","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Palak","family":"Shah","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1940-9706","authenticated-orcid":false,"given":"Gordon","family":"Adams","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John A.","family":"Branda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey D.","family":"Klausner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert","family":"Goldstein","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pardis C.","family":"Sabeti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacob E.","family":"Lemieux","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023053007150567000_2023.05.21.23290304v1.1","doi-asserted-by":"publisher","DOI":"10.1071\/SH19061"},{"key":"2023053007150567000_2023.05.21.23290304v1.2","doi-asserted-by":"publisher","DOI":"10.2471\/BLT.18.228486"},{"issue":"1","key":"2023053007150567000_2023.05.21.23290304v1.3","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.cmi.2022.08.008","article-title":"Global prevalence of Neisseria gonorrhoeae infection in pregnant women: a systematic review and meta-analysis","volume":"29","year":"2023","journal-title":"Clin Microbiol Infect"},{"issue":"1","key":"2023053007150567000_2023.05.21.23290304v1.4","doi-asserted-by":"crossref","first-page":"1152","DOI":"10.1186\/s12879-021-06381-4","article-title":"Gonorrhoea: a systematic review of prevalence reporting globally","volume":"21","year":"2021","journal-title":"BMC Infect Dis"},{"key":"2023053007150567000_2023.05.21.23290304v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.idc.2013.08.004"},{"key":"2023053007150567000_2023.05.21.23290304v1.6","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajog.2016.08.008"},{"key":"2023053007150567000_2023.05.21.23290304v1.7","doi-asserted-by":"crossref","unstructured":"Dolange V , Churchward CP , Christodoulides M , Snyder LAS . 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