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Commonly used laboratory testing methods have many shortcomings. Serological diagnosis has low sensitivity, causing false negatives, while a quantitative real-time polymerase chain reaction (qPCR) requires large equipment and professional staff. To make up for these shortcomings, we proposed a label-free, low-cost, and small-sized ion-sensitive field-effect transistor (ISFET) array based on a low-buffered loop-mediated isothermal amplification (LAMP) assay. A complementary metal oxide semiconductor (CMOS)-based ISFET array with 512 \u00d7 512 sensors was used in this system, which responds specifically to H+ with a sensitivity of 365.7 mV\/pH. For on-chip amplification, a low-buffered LAMP system designed for the conserved sequences of two genes, CARDS and gyrB, was applied. The rapid release of large amounts of H+ in the low-buffered LAMP solution led to a speedy increase in electrical signals captured by the ISFET array, eliminating the need for a sophisticated temperature cycling and optical system. The on-chip results showed that the device can accurately complete MP detection with a detection limit of about 103 copies\/mL (approximately 1 copy per reaction). In the final clinical validation, the detection results of eight throat swab samples using the ISFET sensors were fully consistent with the clinical laboratory diagnostic outcomes, confirming the accuracy and reliability of the ISFET sensors for use in clinical settings. And the entire process from sample lysis to result interpretation takes about 60 min. This platform has potential to be used for the point-of-care testing (POCT) of pathogen infections, providing a basis for the timely adjustment of diagnosis and treatment plans.<\/jats:p>","DOI":"10.3390\/s25051562","type":"journal-article","created":{"date-parts":[[2025,3,4]],"date-time":"2025-03-04T03:26:21Z","timestamp":1741058781000},"page":"1562","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["ISFET Biosensor with Loop-Mediated Isothermal Amplification for Electronic Rapid Detection of Mycoplasma Pneumoniae"],"prefix":"10.3390","volume":"25","author":[{"given":"Jie","family":"Zou","sequence":"first","affiliation":[{"name":"Precision Medicine Translational Research Center (PMTRC), West China Hospital, Sichuan University, Chengdu 610041, China"},{"name":"Department of Laboratory Medicine, Clinical Laboratory Medicine Research Center, West China Hospital, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7516-7639","authenticated-orcid":false,"given":"Jie","family":"Hu","sequence":"additional","affiliation":[{"name":"Precision Medicine Translational Research Center (PMTRC), West China Hospital, Sichuan University, Chengdu 610041, China"},{"name":"Department of Laboratory Medicine, Clinical Laboratory Medicine Research Center, West China Hospital, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Shen","sequence":"additional","affiliation":[{"name":"Precision Medicine Translational Research Center (PMTRC), West China Hospital, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Limei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Precision Medicine Translational Research Center (PMTRC), West China Hospital, Sichuan University, Chengdu 610041, China"},{"name":"Department of Laboratory Medicine, Clinical Laboratory Medicine Research Center, West China Hospital, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weiyi","family":"Bai","sequence":"additional","affiliation":[{"name":"Precision Medicine Translational Research Center (PMTRC), West China Hospital, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"Wang","sequence":"additional","affiliation":[{"name":"One-Chip Biotechnology Co., Ltd., Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianlong","family":"Li","sequence":"additional","affiliation":[{"name":"One-Chip Biotechnology Co., Ltd., Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Yan","sequence":"additional","affiliation":[{"name":"One-Chip Biotechnology Co., Ltd., Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhifeng","family":"Zhang","sequence":"additional","affiliation":[{"name":"One-Chip Biotechnology Co., Ltd., Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0916-5755","authenticated-orcid":false,"given":"Hao","family":"Bai","sequence":"additional","affiliation":[{"name":"Precision Medicine Translational Research Center (PMTRC), West China Hospital, Sichuan University, Chengdu 610041, China"},{"name":"Department of Laboratory Medicine, Clinical Laboratory Medicine Research Center, West China Hospital, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenchuang","family":"Hu","sequence":"additional","affiliation":[{"name":"Precision Medicine Translational Research Center (PMTRC), West China Hospital, Sichuan University, Chengdu 610041, China"},{"name":"Department of Laboratory Medicine, Clinical Laboratory Medicine Research Center, West China Hospital, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Hu, J., Ye, Y., Chen, X., Xiong, L., Xie, W., and Liu, P. 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