Evidence map›Paper›PMID 41165826›Full record

ArticleAnalytical and bioanalytical chemistry2025

Highly sensitive PfAgo-based biosensor for the detection of mutations in the 23S rRNA gene of macrolide-resistant Mycoplasma pneumoniae.

Houqi Wang, Huimin Liao, Huiyun Xie, Suhui He, Xiang Xiao, Haoliang Yin, Zishi Chen, Rong Wei, Zhaoyang Xie, Luxin Yu and 1 more

Abstract read
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Article in Analytical and bioanalytical chemistry, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

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2 · The registry

The trial behind it

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3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Research progress on detection technologies for Mycoplasma pneumoniae.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2026
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4 · The record

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PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

11 authors.

Houqi Wang *Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China.
Huimin Liao *Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China.
Huiyun Xie *Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China.
Suhui HeGuangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China.
Xiang XiaoGuangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China.
Haoliang YinDepartment of Laboratory, The Tenth Affiliated Hospital, Southern Medical University (Dongguan People's Hospital), Dongguan, 523000, China.
Zishi ChenGuangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China.
Rong WeiGuangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China.
Zhaoyang XieGuangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China. xiezhaoyang@gdmu.edu.cn.
Luxin YuGuangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China. yuluxin@gdmu.edu.cn.
Zhangquan ChenGuangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, 523808, China. zqchen@gdmu.edu.cn.ORCID http://orcid.org/0000-0002-1482-046X

Funding

Basic and Applied Basic Research Foundation of Guangdong Province 2023A1515220171Basic and Applied Basic Research Foundation of Guangdong Province 2023A1515220189
6 · The paper itself

Abstract

The prevalence of macrolide-resistant Mycoplasma pneumoniae (M. pneumoniae) is increasing, and rapid macrolide resistance detection assays are needed for clinical management. The A2063G mutation in the 23S rRNA gene of M. pneumoniae is the most common mutation and confers high macrolide resistance. Programmable nucleases have risen as new tools of molecular diagnosis, in which Argonaute is independent of the protospacer adjacent motif (PAM) manner and flexible in molecular diagnosis. By leveraging the specific recognition and cleavage capability of the programmable nuclease Argonaute, we developed a Pyrococcus furiosus Argonaute (PfAgo)-based biosensor for detecting the macrolide resistance-associated A2063G mutation in the 23S rRNA gene of M. pneumoniae without flanking PAM. Through systematic optimization of the PfAgo and guide DNA concentrations to maximize sensitivity and specificity, we achieved a detection limit of 1 fM for DNA single-base mutations, with no cross-reactivity observed against 11 other respiratory tract pathogens. When tested on 25 clinical specimens from patients infected with M. pneumoniae, our assay showed that 20 cases were positive with the A2063G mutation, and 5 cases were negative without the A2063G mutation. Compared to DNA sequencing analysis, our assay demonstrated 100% sensitivity, 100% specificity, and a perfect receiver operating characteristic (area under the curve value = 1.00) (n = 25). We developed a platform for the highly sensitive and specific identification of the macrolide-resistance associated A2063G mutation. This can be used for the detection of the macrolide-resistant M. pneumoniae in clinical specimens and provides a new method for the identification of drug-resistant isolates.

Indexed as

Anti-Bacterial AgentsBiosensing TechniquesDrug Resistance, BacterialMacrolidesMycoplasma pneumoniaeRNA, Ribosomal, 23SHumansLimit of DetectionMutationPneumonia, MycoplasmaAnti-Bacterial AgentsMacrolidesRNA, Ribosomal, 23SBiosensorFluorescence detectionMacrolide resistanceMycoplasma pneumoniaePfAgo

Identifiers

PMID41165826

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.