Multiplex and On-site PCR detection of swine diseases based on the microfluidic chip system

Abstract Background: At present, the process of inspection and quarantine starts with sampling at the customs port, continues with transporting the samples to the central laboratory for inspection experiments, and ends with the inspected results being fed back to the port. This process takes a rather long time, has the risks of degradation of biological samples and generation of pathogenic microorganisms, and does not meet the rapid on-site detection demand. Therefore, development of a technology for rapid and high-throughput detection of pathogenic microorganisms at the customs port is of great significance. This study was to develop a microfluidic chip to be applied to rapid high-throughput detection for swine disease with higher accuracy and lower risk of spreading pathogenic microorganisms during transportation. Results: PCR technology has the advantages of high accurate and sensitivity in disease detection, clinical testing and food quarantine, so it plays an important role in customs inspection. However, the traditional PCR detection instrument has a large size, is time-consuming and has strict requirements on the experimental environment, which greatly limit its application in on-site testing. In this paper, the positive nucleic acid of four swine diseases were detected by a portable and rapid microfluidic PCR system, which could achieve a on-site real-time quantitative PCR detection. Eight clinical samples were detected together on the microfluidic chip in the system, and the detection results were obtained in about an hour. The detection limit of this microfluidic PCR detection system was as low as 1 copies/μL. The results show that the high sensitivity and specificity of the microfluidic PCR detection system in disease detection will play an important role in customs inspection and quarantine during customs clearance. Conclusion: The microfluidic PCR detection system established in this study could meet the requirements for rapid detection of samples at the customs port The new method can avoid the risky process of transporting the samples from the sampling site to the testing lab, and drastically reducing the inspection cycle, and would enable parallel inspections on one chip which greatly raising the efficiency of inspection.

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Multiplex and On-site PCR detection of swine diseases based on the microfluidic chip system

10.21203/rs.2.21078/v1 ◽

2020 ◽

Author(s):

Bin Zhou ◽

Yan Jiang ◽

Yue Wu ◽

Shan Jiang ◽

Kaiming Wang ◽

...

Keyword(s):

High Throughput ◽

Microfluidic Chip ◽

Detection System ◽

Sampling Site ◽

Pcr Detection ◽

Clinical Samples ◽

Disease Detection ◽

Pathogenic Microorganisms ◽

Inspection And Quarantine ◽

Swine Diseases

Abstract Background：At present, the process of inspection and quarantine starts with sampling at the customs port, continues with transporting the samples to the central laboratory for inspection experiments, and ends with the inspected results being fed back to the port. This process takes a rather long time, has the risks of degradation of biological samples [32] and generation of pathogenic microorganisms [33], and does not meet the rapid on-site detection demand [34]. Therefore, development of a technology for rapid and high- throughput detection of pathogenic microorganisms at the customs port is of great significance. This study was to develop a microfluidic chip to be applied to rapid high-throughput swine disease detection with higher accuracy and lower risk of spreading pathogenic microorganisms during transportation.Results: PCR technology has the advantages of high accurate and sensitivity in disease detection, clinical testing and food quarantine, so it plays an important role in customs inspection. However, the traditional PCR detection instrument has a large size, is time-consuming and has strict requirements on the experimental environment, which greatly limit its application in on-site testing. In this paper, the clinical samples of four swine diseases were detected by a portable and rapid microfluidic PCR system, which could achieve a on- site real-time quantitative PCR detection. Eight clinical samples were detected together on the microfluidic chip in the system, and the detection results were obtained in about an hour. The detection limit of this microfluidic PCR detection system was as low as 1 copies/μL. The results show that the high sensitivity and specificity of the microfluidic PCR detection system in disease detection will play an important role in customs inspection and quarantine during customs clearance.Conclusion: The microfluidic PCR detection system established in this study could meet the requirements for rapid detection of samples at the customs portThe new method can avoid the risky process of transporting the samples from the sampling site to the testing lab, and drastically reducing the inspection cycle, and would enable parallel inspections on one chip which greatly raising the efficiency of inspection.

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Multiplex and On-site PCR detection of swine diseases based on the microfluidic chip system

10.21203/rs.2.21078/v3 ◽

2020 ◽

Author(s):

Yan Jiang ◽

Shan Jiang ◽

Yue Wu ◽

Bin Zhou ◽

Kaiming Wang ◽

...

Keyword(s):

High Throughput ◽

Microfluidic Chip ◽

Detection System ◽

Sampling Site ◽

High Sensitivity ◽

Pcr Detection ◽

Clinical Samples ◽

Pathogenic Microorganisms ◽

Real Time Quantitative Pcr ◽

Inspection And Quarantine

Abstract Background： At present, the process of inspection and quarantine starts with sampling at the customs port, continues with transporting the samples to the central laboratory for inspection experiments, and ends with the inspected results being fed back to the port. This process had the risks of degradation of biological samples and generation of pathogenic microorganisms and did not meet the rapid on-site detection demand because it took a rather long time. Therefore, it is urgently needed to develop a rapid and high-throughput detection assay of pathogenic microorganisms at the customs port. The aim of this study was to develop a microfluidic chip to rapidly detect swine pathogenic microorganisms with high-throughput and higher accuracy. Moreover, this chip will decrease the risk of spreading infection during transportation.Results: A series of experiments were performed to establish a microfluidic chip. The resulting data showed that the positive nucleic acid of four swine viruses were detected by using a portable and rapid microfluidic PCR system, which could achieve a on-site real-time quantitative PCR detection. Furthermore, the detection results of eight clinical samples were obtained within an hour. The detection limit of this microfluidic PCR detection system was as low as 1 copies/μL. The results showed that the high sensitivity and specificity of this chip system in disease detection played an important role in customs inspection and quarantine during customs clearance.Conclusion: The microfluidic PCR detection system established in this study could meet the requirement for rapid detection of samples at the customs port. This chip could avoid the risky process of transporting the samples from the sampling site to the testing lab, and drastically reduce the inspection cycle. Moreover, it would enable parallel inspections on one chip, which greatly raised the efficiency of inspection.

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Multiplex and On-site PCR detection of swine diseases based on the microfluidic chip system

10.21203/rs.2.21078/v6 ◽

2021 ◽

Author(s):

Yan Jiang ◽

Shan Jiang ◽

Yue Wu ◽

Bin Zhou ◽

Kaiming Wang ◽

...

Keyword(s):

High Throughput ◽

Microfluidic Chip ◽

Detection System ◽

Sampling Site ◽

High Sensitivity ◽

Pcr Detection ◽

Clinical Samples ◽

Pathogenic Microorganisms ◽

Real Time Quantitative Pcr ◽

Inspection And Quarantine

Abstract Background: At present, the process of inspection and quarantine starts with sampling at the customs port, continues with transporting the samples to the central laboratory for inspection experiments, and ends with the inspected results being fed back to the port. This process had the risks of degradation of biological samples and generation of pathogenic microorganisms and did not meet the rapid on-site detection demand because it took a rather long time. Therefore, it is urgently needed to develop a rapid and high-throughput detection assay of pathogenic microorganisms at the customs port. The aim of this study was to develop a microfluidic chip to rapidly detect swine pathogenic microorganisms with high-throughput and higher accuracy. Moreover, this chip will decrease the risk of spreading infection during transportation.Results: A series of experiments were performed to establish a microfluidic chip. The resulting data showed that the positive nucleic acid of four swine viruses were detected by using a portable and rapid microfluidic PCR system, which could achieve a on-site real-time quantitative PCR detection. Furthermore, the detection results of eight clinical samples were obtained within an hour. The detection limit of this microfluidic PCR detection system was as low as 1 copies/μL. The results showed that the high sensitivity and specificity of this chip system in disease detection played an important role in customs inspection and quarantine during customs clearance.Conclusion: The microfluidic PCR detection system established in this study could meet the requirement for rapid detection of samples at the customs port. This chip could avoid the risky process of transporting the samples from the sampling site to the testing lab, and drastically reduce the inspection cycle. Moreover, it would enable parallel inspections on one chip, which greatly raised the efficiency of inspection.

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Multiplex and On-site PCR detection of swine diseases based on the microfluidic chip system

10.21203/rs.2.21078/v5 ◽

2020 ◽

Author(s):

Yan Jiang ◽

Shan Jiang ◽

Yue Wu ◽

Bin Zhou ◽

Kaiming Wang ◽

...

Keyword(s):

High Throughput ◽

Microfluidic Chip ◽

Detection System ◽

Sampling Site ◽

High Sensitivity ◽

Pcr Detection ◽

Clinical Samples ◽

Pathogenic Microorganisms ◽

Real Time Quantitative Pcr ◽

Inspection And Quarantine

Abstract Background： At present, the process of inspection and quarantine starts with sampling at the customs port, continues with transporting the samples to the central laboratory for inspection experiments, and ends with the inspected results being fed back to the port. This process had the risks of degradation of biological samples and generation of pathogenic microorganisms and did not meet the rapid on-site detection demand because it took a rather long time. Therefore, it is urgently needed to develop a rapid and high-throughput detection assay of pathogenic microorganisms at the customs port. The aim of this study was to develop a microfluidic chip to rapidly detect swine pathogenic microorganisms with high-throughput and higher accuracy. Moreover, this chip will decrease the risk of spreading infection during transportation.Results: A series of experiments were performed to establish a microfluidic chip. The resulting data showed that the positive nucleic acid of four swine viruses were detected by using a portable and rapid microfluidic PCR system, which could achieve a on-site real-time quantitative PCR detection. Furthermore, the detection results of eight clinical samples were obtained within an hour. The detection limit of this microfluidic PCR detection system was as low as 1 copies/μL. The results showed that the high sensitivity and specificity of this chip system in disease detection played an important role in customs inspection and quarantine during customs clearance.Conclusion: The microfluidic PCR detection system established in this study could meet the requirement for rapid detection of samples at the customs port. This chip could avoid the risky process of transporting the samples from the sampling site to the testing lab, and drastically reduce the inspection cycle. Moreover, it would enable parallel inspections on one chip, which greatly raised the efficiency of inspection.

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Multiplex and on-site PCR detection of swine diseases based on the microfluidic chip system

BMC Veterinary Research ◽

10.1186/s12917-021-02825-w ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Yan Jiang ◽

Shan Jiang ◽

Yue Wu ◽

Bin Zhou ◽

Kaimin Wang ◽

...

Keyword(s):

High Throughput ◽

Microfluidic Chip ◽

Detection System ◽

Sampling Site ◽

High Specificity ◽

Pcr Detection ◽

Clinical Samples ◽

Pathogenic Microorganisms ◽

Real Time Quantitative Pcr ◽

Inspection And Quarantine

Abstract Background At present, the process of inspection and quarantine starts with sampling at the customs port, continues with transporting the samples to the central laboratory for inspection experiments, and ends with the inspected results being fed back to the port. This process had the risks of degradation of biological samples and generation of pathogenic microorganisms and did not meet the rapid on-site detection demand because it took a rather long time. Therefore, it is urgently needed to develop a rapid and high-throughput detection assay of pathogenic microorganisms at the customs port. The aim of this study was to develop a microfluidic chip to rapidly detect swine pathogenic microorganisms with high-throughput and higher accuracy. Moreover, this chip will decrease the risk of spreading infection during transportation. Results A series of experiments were performed to establish a microfluidic chip. The resulting data showed that the positive nucleic acid of four swine viruses were detected by using a portable and rapid microfluidic PCR system, which could achieve a on-site real-time quantitative PCR detection. Furthermore, the detection results of eight clinical samples were obtained within an hour. The lowest concentration that amplified of this microfluidic PCR detection system was as low as 1 copies/μL. The results showed that the high specificity of this chip system in disease detection played an important role in customs inspection and quarantine during customs clearance. Conclusion The microfluidic PCR detection system established in this study could meet the requirement for rapid detection of samples at the customs port. This chip could avoid the risky process of transporting the samples from the sampling site to the testing lab, and drastically reduce the inspection cycle. Moreover, it would enable parallel inspections on one chip, which greatly raised the efficiency of inspection.

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Multiplex and On-site PCR detection of swine diseases based on the microfluidic chip system

10.21203/rs.2.21078/v4 ◽

2020 ◽

Author(s):

Yan Jiang ◽

Shan Jiang ◽

Yue Wu ◽

Bin Zhou ◽

Kaiming Wang ◽

...

Keyword(s):

High Throughput ◽

Microfluidic Chip ◽

Detection System ◽

Sampling Site ◽

High Sensitivity ◽

Pcr Detection ◽

Clinical Samples ◽

Pathogenic Microorganisms ◽

Real Time Quantitative Pcr ◽

Inspection And Quarantine

Abstract Background： At present, the process of inspection and quarantine starts with sampling at the customs port, continues with transporting the samples to the central laboratory for inspection experiments, and ends with the inspected results being fed back to the port. This process had the risks of degradation of biological samples and generation of pathogenic microorganisms and did not meet the rapid on-site detection demand because it took a rather long time. Therefore, it is urgently needed to develop a rapid and high-throughput detection assay of pathogenic microorganisms at the customs port. The aim of this study was to develop a microfluidic chip to rapidly detect swine pathogenic microorganisms with high-throughput and higher accuracy. Moreover, this chip will decrease the risk of spreading infection during transportation. Results: A series of experiments were performed to establish a microfluidic chip. The resulting data showed that the positive nucleic acid of four swine viruses were detected by using a portable and rapid microfluidic PCR system, which could achieve a on-site real-time quantitative PCR detection. Furthermore, the detection results of eight clinical samples were obtained within an hour. The detection limit of this microfluidic PCR detection system was as low as 1 copies/μL. The results showed that the high sensitivity and specificity of this chip system in disease detection played an important role in customs inspection and quarantine during customs clearance. Conclusion: The microfluidic PCR detection system established in this study could meet the requirement for rapid detection of samples at the customs port. This chip could avoid the risky process of transporting the samples from the sampling site to the testing lab, and drastically reduce the inspection cycle. Moreover, it would enable parallel inspections on one chip, which greatly raised the efficiency of inspection.

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Miniaturized high throughput detection system for capillary array electrophoresis on chip with integrated light emitting diode array as addressed ring-shaped light source

Lab on a Chip ◽

10.1039/b810705j ◽

2009 ◽

Vol 9 (5) ◽

pp. 733-736 ◽

Cited By ~ 18

Author(s):

Kangning Ren ◽

Qionglin Liang ◽

Xuan Mu ◽

Guoan Luo ◽

Yiming Wang

Keyword(s):

Light Source ◽

High Throughput ◽

Detection System ◽

Light Emitting Diode ◽

Diode Array ◽

Light Emitting ◽

Capillary Array Electrophoresis ◽

High Throughput Detection ◽

Capillary Array ◽

On Chip

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The “TSH Receptor Glo Assay” – A High-Throughput Detection System for Thyroid Stimulation

Frontiers in Endocrinology ◽

10.3389/fendo.2016.00003 ◽

2016 ◽

Vol 7 ◽

Cited By ~ 6

Author(s):

Rauf Latif ◽

Zerlina Lau ◽

Pamela Cheung ◽

Dan P. Felsenfeld ◽

Terry F. Davies

Keyword(s):

High Throughput ◽

Detection System ◽

Tsh Receptor ◽

High Throughput Detection

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High throughput detection and genetic epidemiology of SARS-CoV-2 using COVIDSeq next-generation sequencing

PLoS ONE ◽

10.1371/journal.pone.0247115 ◽

2021 ◽

Vol 16 (2) ◽

pp. e0247115

Author(s):

Rahul C. Bhoyar ◽

Abhinav Jain ◽

Paras Sehgal ◽

Mohit Kumar Divakar ◽

Disha Sharma ◽

...

Keyword(s):

High Throughput ◽

Genetic Epidemiology ◽

Confirmatory Test ◽

Clinical Samples ◽

Rt Pcr ◽

Additional Advantage ◽

Depth Analysis ◽

High Concordance ◽

High Throughput Detection ◽

First Time

The rapid emergence of coronavirus disease 2019 (COVID-19) as a global pandemic affecting millions of individuals globally has necessitated sensitive and high-throughput approaches for the diagnosis, surveillance, and determining the genetic epidemiology of SARS-CoV-2. In the present study, we used the COVIDSeq protocol, which involves multiplex-PCR, barcoding, and sequencing of samples for high-throughput detection and deciphering the genetic epidemiology of SARS-CoV-2. We used the approach on 752 clinical samples in duplicates, amounting to a total of 1536 samples which could be sequenced on a single S4 sequencing flow cell on NovaSeq 6000. Our analysis suggests a high concordance between technical duplicates and a high concordance of detection of SARS-CoV-2 between the COVIDSeq as well as RT-PCR approaches. An in-depth analysis revealed a total of six samples in which COVIDSeq detected SARS-CoV-2 in high confidence which were negative in RT-PCR. Additionally, the assay could detect SARS-CoV-2 in 21 samples and 16 samples which were classified inconclusive and pan-sarbeco positive respectively suggesting that COVIDSeq could be used as a confirmatory test. The sequencing approach also enabled insights into the evolution and genetic epidemiology of the SARS-CoV-2 samples. The samples were classified into a total of 3 clades. This study reports two lineages B.1.112 and B.1.99 for the first time in India. This study also revealed 1,143 unique single nucleotide variants and added a total of 73 novel variants identified for the first time. To the best of our knowledge, this is the first report of the COVIDSeq approach for detection and genetic epidemiology of SARS-CoV-2. Our analysis suggests that COVIDSeq could be a potential high sensitivity assay for the detection of SARS-CoV-2, with an additional advantage of enabling the genetic epidemiology of SARS-CoV-2.

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