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Updated: Dec 11, 2025
High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR
Published on: November 28, 2016
Establishment and application of a multiple cross displacement amplification combined with nanoparticles-based
Shijun Li1, Chunting Liu1, Ying Liu1
1Laboratory of Bacterial Infectious Disease of Experimental Center, Guizhou Provincial Center for Disease Control and Prevention, Guiyang, 550004, Guizhou, China.
Background:
Bordetella pertussis is the causative agent of pertussis, a respiratory tract infectious disease. Efficient techniques for detection of B. pertussis isolates are important for clinical diagnosis. Multiple cross displacement amplification (MCDA), a novel isothermal amplification based molecular detection method, has been developed to overcome the technical drawback of the current methods in recent years. This aim of this study is to develop a MCDA with Nanoparticles-based Lateral Flow Biosensor (MCDA-LFB) for the detection of B. pertussis.
Results:
A set of 10 primers based on the pertussis toxin (PT) promoter region sequence of B. pertussis was designed. The B. pertussis-MCDA-LFB assay was successfully established and optimized at 64 °C for reaction of 40 min. The detection limit was determined as 10 fg/reaction of pure DNA, and no cross-reactions to non-B. pertussis strains were observed, based on the specificity validation. The whole operation, ranging from template preparation to result reporting, could be completed within 70 min without requirement of costly equipment. The B. pertussis-MCDA-LFB in clinic sample detection yielded identical positive rates with traditional culture and showed higher sensitivity than conventional PCR. The results of MCDA-LFB are easier to read due to the usage of LFB.
Conclusions:
The isothermal amplification based MCDA-LFB established in the present study is a specific, sensitive, rapid and economical technique for the detection of B. pertussis.
Insights
A new method called Multiple Cross Displacement Amplification with Nanoparticles-based Lateral Flow Biosensor (MCDA-LFB) offers a rapid and sensitive way to detect Bordetella pertussis. This technique is accurate for clinical diagnosis and overcomes limitations of current detection methods.
Area of Science:
- Microbiology
- Molecular Biology
- Biotechnology
Background:
- Bordetella pertussis causes the infectious respiratory disease pertussis.
- Accurate detection of B. pertussis is crucial for clinical diagnosis.
- Multiple Cross Displacement Amplification (MCDA) is a novel isothermal amplification method addressing limitations of existing techniques.
Purpose of the Study:
- To develop a MCDA combined with a Nanoparticles-based Lateral Flow Biosensor (MCDA-LFB) for detecting B. pertussis.
Main Methods:
- Designed 10 primers targeting the pertussis toxin (PT) promoter region of B. pertussis.
- Optimized the B. pertussis-MCDA-LFB assay at 64°C for 40 minutes.
- Validated specificity against non-B. pertussis strains.
Main Results:
- Achieved a detection limit of 10 fg/reaction for pure DNA with high specificity.
- The entire process, from sample to result, was completed within 70 minutes without specialized equipment.
- Clinical sample detection showed identical positive rates to traditional culture and higher sensitivity than conventional PCR.
- Lateral Flow Biosensor (LFB) facilitated easier result interpretation.
Conclusions:
- The developed MCDA-LFB is a specific, sensitive, rapid, and cost-effective method for B. pertussis detection.
- This isothermal amplification-based technique offers a valuable tool for clinical diagnosis.
