---
title: "Ultrasensitive detection of Mycobacterium tuberculosis rpoB 531 mutation using MCC‑PPA"
id: "pubmed-42764695"
canonical_url: "https://medichelpline.com/clinical-feed/pubmed-42764695"
content_type: "clinical_feed_article"
specialty: "Infectious Disease"
source_name: "PubMed / NCBI"
source_url: "https://pubmed.ncbi.nlm.nih.gov/42764695/"
doi: "10.1039/d6an00774k"
published_at: "2026-09-21T00:00:00.000Z"
evidence_level: "Journal Article"
license: "CC-BY-NC-4.0 / Informational Use"
---
# Ultrasensitive detection of Mycobacterium tuberculosis rpoB 531 mutation using MCC‑PPA
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/pubmed-42764695
- **Specialty:** [Infectious Disease](https://medichelpline.com/clinical-feed/infectious-disease.md)
- **Primary Source:** PubMed / NCBI
- **Source URL:** [Original Journal Publication](https://pubmed.ncbi.nlm.nih.gov/42764695/)
- **DOI:** [10.1039/d6an00774k](https://doi.org/10.1039%2Fd6an00774k)
- **Published At:** 2026-09-21T00:00:00.000Z
- **Evidence Rating:** Journal Article
## Executive GIST (TL;DR)
- The authors developed a novel assay named **MCC‑PPA** (molecular crowding conditions combined with padlock probe amplification) to detect the rpoB 531 (TCG > TTG) mutation in **Mycobacterium tuberculosis**, which is associated with **rifampicin resistance**. - **Polyethylene glycol (PEG)** was used to create molecular crowding during padlock probe ligation, improving probe cyclization and subsequent amplification efficiency. - The assay demonstrated a wide linear dynamic range from **10 aM to 1 nM** and a reported lower limit of detection of **4.52 aM**, with a total assay time of **1.5 hours**. - Sensitivity was improved **400‑fold** compared with padlock probe amplification conducted without PEG (PEG‑free assay). - Clinical evaluation included **105 clinical specimens**: 21 with the rpoB 531 mutation and 84 without. The MCC‑PPA assay achieved **100% sensitivity** and **100% specificity** when benchmarked against DNA sequencing. - Compared with fluorescence PCR melting curve analysis, the MCC‑PPA assay also showed **100% clinical sensitivity** for detecting the rpoB 531 TCG > TTG mutation. - The findings indicate strong applicability of MCC‑PPA for detecting antimicrobial resistance mutations in **M. tuberculosis**, providing a rapid and highly sensitive molecular diagnostic option. - Details such as exact experimental conditions, primer/probe sequences, sample preparation workflow, and broader validation cohorts were not reported in the abstract and would require consulting the full article for implementation specifics.
## Clinical Analysis & Structured Key Points
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Online ahead of print. # Ultrasensitive detection of drug resistance genes in _Mycobacterium tuberculosis_ based on molecular crowding conditions coupled with padlock probe amplification [Miaofen Lin](https://pubmed.ncbi.nlm.nih.gov/?term=Lin+M&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#full-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn.")[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#full-view-affiliation-2 "Department of Clinical Laboratory, Centre for Chronic Disease Control and Prevention of Yunfu, Yunfu, China."), [Jing He](https://pubmed.ncbi.nlm.nih.gov/?term=He+J&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#full-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Jiaxin Wang](https://pubmed.ncbi.nlm.nih.gov/?term=Wang+J&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#full-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Huiqin Zhu](https://pubmed.ncbi.nlm.nih.gov/?term=Zhu+H&cauthor_id=42764695)[ 3 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#full-view-affiliation-3 "Department of Clinical Laboratory, YunFu People's Hospital, Yunfu, China."), [Hengming Ye](https://pubmed.ncbi.nlm.nih.gov/?term=Ye+H&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#full-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Suhui He](https://pubmed.ncbi.nlm.nih.gov/?term=He+S&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#full-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Weilong Zhang](https://pubmed.ncbi.nlm.nih.gov/?term=Zhang+W&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#full-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Luxin Yu](https://pubmed.ncbi.nlm.nih.gov/?term=Yu+L&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#full-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Zhangquan Chen](https://pubmed.ncbi.nlm.nih.gov/?term=Chen+Z&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#full-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn.") Affiliations Expand ### Affiliations * 1 Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn. * 2 Department of Clinical Laboratory, Centre for Chronic Disease Control and Prevention of Yunfu, Yunfu, China. * 3 Department of Clinical Laboratory, YunFu People's Hospital, Yunfu, China. * PMID: **42764695** * DOI: [ 10.1039/d6an00774k ](https://doi.org/10.1039/d6an00774k) Item in Clipboard # Ultrasensitive detection of drug resistance genes in _Mycobacterium tuberculosis_ based on molecular crowding conditions coupled with padlock probe amplification Miaofen Lin et al. Analyst. 2026. Show details Display options Display options Format Abstract PubMed PMID Analyst Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Analyst%22%5Bjour%5D&sort=date&sort_order=desc) * [ Search in NLM Catalog ](https://www.ncbi.nlm.nih.gov/nlmcatalog?term=%22Analyst%22%5BTitle+Abbreviation%5D) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42764695/) . 2026 Sep 21. doi: 10.1039/d6an00774k. Online ahead of print. ### Authors [Miaofen Lin](https://pubmed.ncbi.nlm.nih.gov/?term=Lin+M&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#short-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn.")[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#short-view-affiliation-2 "Department of Clinical Laboratory, Centre for Chronic Disease Control and Prevention of Yunfu, Yunfu, China."), [Jing He](https://pubmed.ncbi.nlm.nih.gov/?term=He+J&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#short-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Jiaxin Wang](https://pubmed.ncbi.nlm.nih.gov/?term=Wang+J&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#short-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Huiqin Zhu](https://pubmed.ncbi.nlm.nih.gov/?term=Zhu+H&cauthor_id=42764695)[ 3 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#short-view-affiliation-3 "Department of Clinical Laboratory, YunFu People's Hospital, Yunfu, China."), [Hengming Ye](https://pubmed.ncbi.nlm.nih.gov/?term=Ye+H&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#short-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Suhui He](https://pubmed.ncbi.nlm.nih.gov/?term=He+S&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#short-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Weilong Zhang](https://pubmed.ncbi.nlm.nih.gov/?term=Zhang+W&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#short-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Luxin Yu](https://pubmed.ncbi.nlm.nih.gov/?term=Yu+L&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#short-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn."), [Zhangquan Chen](https://pubmed.ncbi.nlm.nih.gov/?term=Chen+Z&cauthor_id=42764695)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42764695/#short-view-affiliation-1 "Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn.") ### Affiliations * 1 Guangdong Provincial Key Laboratory of Medical Immunology and Molecular Diagnostics, The First Dongguan Affiliated Hospital, School of Medical Technology, Guangdong Medical University, Dongguan, China. yuluxin@gdmu.edu.cn. * 2 Department of Clinical Laboratory, Centre for Chronic Disease Control and Prevention of Yunfu, Yunfu, China. * 3 Department of Clinical Laboratory, YunFu People's Hospital, Yunfu, China. * PMID: **42764695** * DOI: [ 10.1039/d6an00774k ](https://doi.org/10.1039/d6an00774k) Item in Clipboard Cite Display options Display options Format Abstract PubMed PMID ## Abstract In this work, we developed a novel approach driven by molecular crowding conditions combined with padlock probe amplification (MCC-PPA) to detect the _rpoB 531_ (TCG > TTG) mutation in _Mycobacterium tuberculosis_ associated with rifampicin resistance. Polyethylene glycol (PEG) was employed to create molecular crowding conditions in the ligation of the padlock probe, enhancing padlock probe cyclization and increasing the amplification efficiency. This proposed assay displayed a broader linear range (10 aM-1 nM) and a lower detection limit (4.52 aM) within 1.5 hours. The sensitivity of this assay was enhanced 400-fold over the PEG-free padlock probe amplification assay. When evaluated on 105 clinical specimens, including 21 with the _rpoB 531_ mutation and 84 without this mutation, the assay demonstrated 100% sensitivity and 100% specificity relative to DNA sequencing results. When compared against fluorescence PCR melting curve analysis, the clinical sensitivity for detecting _Mycobacterium tuberculosis_ rifampicin resistance gene _rpoB 531_ TCG > TTG mutation was also 100%. These findings highlight our proposed assay's strong applicability for the detection of antimicrobial resistance mutations in _Mycobacterium tuberculosis_. 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