Amino-modified silica membrane capable of DNA extraction and enrichment for facilitated isothermal amplification detection of Mycoplasma pneumoniae

Herein, we developed a facile integrated Mycoplasma pneumoniae diagnosis platform by combining amino-modified silica membrane (AMSM)-based nucleic acids fast extraction and enrichment with colorimetric isothermal amplification detection. AMSM demonstrates a strong ability to capture and enrich nucle...

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Veröffentlicht in:Journal of pharmaceutical and biomedical analysis 2023-02, Vol.224, p.115190-115190, Article 115190
Hauptverfasser: Duan, Yake, Zhang, Xin, Li, Yong, Zhao, Xiaoli, Zhao, Xiaowen, Chen, Lei, Shi, Chao, Ma, Cuiping, Wang, Xiujuan
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Sprache:eng
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Zusammenfassung:Herein, we developed a facile integrated Mycoplasma pneumoniae diagnosis platform by combining amino-modified silica membrane (AMSM)-based nucleic acids fast extraction and enrichment with colorimetric isothermal amplification detection. AMSM demonstrates a strong ability to capture and enrich nucleic acids in complicated biological matrices, and the purified AMSM/nucleic acids composite could be directly used to perform isothermal amplification including denaturation bubble-mediated strand exchange amplification (SEA) and loop-mediated isothermal amplification (LAMP) reactions. Through comparing clinical specimens, excellent performance of AMSM-based SEA assay with 93.33% sensitivity and 100% specificity relative to real-time PCR was observed, and for AMSM-based LAMP was 96.67% and 100%, respectively. The diagnostic procedure could be completed within 55 min, and the colorimetric-based visual result further alleviates the use of sophisticated equipment. The proposed approach possesses great potential as a simple and time-saving alternative for point-of-care testing (POCT) of M. pneumoniae in resource-limited regions. [Display omitted] •AMSM-based nucleic acid extraction with colorimetric isothermal amplification for M. pneumoniae detection was established.•AMSM allows the fast extraction and enrichment of nucleic acids within 3 min.•AMSM has the potential to be used for analyzing low-concentration samples.•The AMSM/nucleic acids composite, without nucleic acid elution step, could be directly used to perform amplification.•The result was observed directly by the naked eyes through color change.
ISSN:0731-7085
1873-264X
DOI:10.1016/j.jpba.2022.115190