Rapid detection of multiple pathogens by the combined loop-mediated isothermal amplification technology and microfluidic chip technology

Ou, Hongling; Wang, Yan; Wang, Qiyuan; Ma, Yingying; Liu, Chen; Jia, Lianling; Zhang, Qiaoyun; Li, Mengwen; Feng, Xinxin; Li, Meng; Wang, Xinru; Wang, Chengbin

doi:10.21037/apm-21-2792

Cited by 5 publications

(6 citation statements)

References 18 publications

(18 reference statements)

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“… 17 At the same time, the LAMP microfluidic chip is divided into multiple reaction units, which can parallel detect a variety of pathogens and ensure the independence of each reaction and the accuracy of the results. 27 Finally, the method also has the characteristics of user‐friendliness and simpleness. The system is small and portable, does not require professionals to operate, and is more suitable for under‐resourced areas where often no resources for special training for medical staff.…”

Section: Discussionmentioning

confidence: 99%

“…Not only eliminates the need for DNA extraction, avoids possible contamination during the extraction process and the loss of a large amount DNA in the sample but also saves a lot of the time of preprocessing and shortens the entire detection cycle 17 . At the same time, the LAMP microfluidic chip is divided into multiple reaction units, which can parallel detect a variety of pathogens and ensure the independence of each reaction and the accuracy of the results 27 . Finally, the method also has the characteristics of user‐friendliness and simpleness.…”

Section: Discussionmentioning

confidence: 99%

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Comparative analysis of loop‐mediated isothermal amplification combined with microfluidic chip technology and q‐PCR in the detection of clinical infectious pathogens

Zhang

Ou²,

Jia³

et al. 2022

Clinical Laboratory Analysis

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Background Rapid diagnosis of infectious pathogens at an early stage is crucial to stabilize the patient's condition, reduce medical costs, and shorten hospital stays. Currently, some point‐of‐care tests have their own shortcomings. Therefore, we built a microfluidic chip based on loop‐mediated isothermal amplification to can quickly and sensitively detect infectious pathogens. Methods We extracted the DNA of S. aureus, MRSA, Shigella and Klebsiella pneumoniae. Then, the DNA samples were diluted by 10‐fold and examined by two methods: LAMP‐microfluidic chip and q‐PCR, the sensitivity of whom was also compared. In addition, the specificity of the two was also examined by detecting the target bacteria and other microorganisms using the same methods. Finally, we extracted and tested the DNA of clinically infected humoral samples to determine the coincidence rate between the two methods and the bacterial culture method. Results For S. aureus, MRSA, Shigella, and Klebsiella pneumoniae, the detection limits of the chip were 2.25 × 103 copies/μl, 5.32 × 103 copies/μl, 2.89 × 103 copies/μl, 6.53 × 102 copies/μl, and the detection limits of q‐PCR were 2.25 × 102 copies/μl, 5.32 × 101 copies/μl, 2.89 × 102 copies/μl, 6.53 × 101 copies/μl, respectively. In terms of detection specificity, neither method cross‐reacted with other strains. For the detection of infectious humoral samples, the total coincidence rate between the q‐PCR and bacterial culture method was 85.7%, 95%, 95%, and 95.5%, and the total coincidence rate between the chip and bacterial culture method was 81%, 95%, 90%, and 86.4%, respectively. Conclusion LAMP‐microfluidic chip provides a simple, sensitive, specific, convenient, and rapid pathogen detection method for clinically infected humoral samples without relying on expensive equipment or technical personnels.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Comparative analysis of loop‐mediated isothermal amplification combined with microfluidic chip technology and q‐PCR in the detection of clinical infectious pathogens

Zhang

Ou²,

Jia³

et al. 2022

Clinical Laboratory Analysis

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“…Currently, LAMP technology on LOC has been applied to the detection of breast cancer biomarkers, dengue serotypes, carbapenemase-producing organisms, and multiple pathogens in the lower reproductive tract during pregnancy (Moser et al, 2022;Wormald et al, 2022;Wu et al, 2022;Xu et al, 2022). Microfluidic chips are characterized by high throughput and low cost, a method of LAMP technology combined with a microfluidic chip to detect a Li et al 10.3389/fcimb.2022.1068015 variety of pathogens has been established, which can perform detection of multiple pathogens and real-time detection within 40 min without aerosol pollution and cross reaction (Ou et al, 2021). Research on the combined application of LAMP technology and microfluidic chips to detect pathogens is of great significance for the rapid detection of SARS-CoV-2 infection.…”

Section: Lamp Combined With Microfluidic Technologymentioning

confidence: 99%

A loop-mediated isothermal amplification-enabled analytical assay for the detection of SARS-CoV-2: A review

Sun

et al. 2022

Front. Cell. Infect. Microbiol.

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The number of words: 4645, the number of figures: 4, the number of tables: 1The outbreak of COVID-19 in December 2019 caused a global pandemic of acute respiratory disease, and with the increasing virulence of mutant strains and the number of confirmed cases, this has resulted in a tremendous threat to global public health. Therefore, an accurate diagnosis of COVID-19 is urgently needed for rapid control of SARS-CoV-2 transmission. As a new molecular biology technology, loop-mediated isothermal amplification (LAMP) has the advantages of convenient operation, speed, low cost and high sensitivity and specificity. In the past two years, rampant COVID-19 and the continuous variation in the virus strains have demanded higher requirements for the rapid detection of pathogens. Compared with conventional RT–PCR and real-time RT–PCR methods, genotyping RT-LAMP method and LAMP plus peptide nucleic acid (PNA) probe detection methods have been developed to correctly identified SARS-CoV-2 variants, which is also why LAMP technology has attracted much attention. LAMP detection technology combined with lateral flow assay, microfluidic technology and other sensing technologies can effectively enhance signals by nucleic acid amplification and help to give the resulting output in a faster, more convenient and user-friendly way. At present, LAMP plays an important role in the detection of SARS-CoV-2.

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“…There has been some work using LAMP to identify Candida species in an isothermal and multi-target way [28][29][30][31][32]. Some works also realized the detection with the participation of microfluidic chips [33,34]. However, none of them were involved in identifying different Candida species in clinical samples of VVC, especially C. glabrata, C. parapsilosis, and C. tropicalis.…”

Section: Introductionmentioning

confidence: 99%

An Integrated and Multi-Target Nucleic Acid Isothermal Analysis System for Rapid Diagnosis of Vulvovaginal Candidiasis

Jin

Mao

et al. 2023

Biosensors

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Rapid identification of Candida species is significant for the diagnosis of vulvovaginal candidiasis (VVC). An integrated and multi-target system for the rapid, high-specificity, and high-sensitivity detection of four Candida species was developed. The system consists of a rapid sample processing cassette and a rapid nucleic acid analysis device. The cassette could process the Candida species to release nucleic acids in 15 min. The released nucleic acids were analyzed by the device as fast as within 30 min, using the loop-mediated isothermal amplification method. The four Candida species could be simultaneously identified, with each reaction using only 1.41 µL of reaction mixture, which was low cost. The RPT (rapid sample processing and testing) system could detect the four Candida species with high sensitivity (<2 CFU/reaction) and high specificity. The system also processed and analyzed 32 clinical samples, giving the results with high clinical sensitivity and specificity. Hence, the system was a significant and effective platform for the diagnosis of VVC. Furthermore, the period of validity of the reagents and chips used in the system was >90 days, and the system could also be used for the detection of bacteria.

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Rapid detection of multiple pathogens by the combined loop-mediated isothermal amplification technology and microfluidic chip technology

Cited by 5 publications

References 18 publications

Comparative analysis of loop‐mediated isothermal amplification combined with microfluidic chip technology and q‐PCR in the detection of clinical infectious pathogens

Comparative analysis of loop‐mediated isothermal amplification combined with microfluidic chip technology and q‐PCR in the detection of clinical infectious pathogens

A loop-mediated isothermal amplification-enabled analytical assay for the detection of SARS-CoV-2: A review

An Integrated and Multi-Target Nucleic Acid Isothermal Analysis System for Rapid Diagnosis of Vulvovaginal Candidiasis

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