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

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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

2020 ◽  
Author(s):  
Yan Jiang ◽  
Shan Jiang ◽  
Yue Wu ◽  
Bin Zhou ◽  
Kaiming Wang ◽  
...  
Keyword(s):  
High Throughput ◽  
Microfluidic Chip ◽  
Detection System ◽  
Pcr Detection ◽  
Clinical Samples ◽  
Disease Detection ◽  
Pathogenic Microorganisms ◽  
Inspection And Quarantine ◽  
High Throughput Detection ◽  
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 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.

scholarly journals High-throughput HTLV-1 proviral DNA detection system using a nucleic acid extraction robot and real-time PCR detection.

2001 ◽  
Vol 47 (3) ◽  
pp. 378-383 ◽  
Author(s):  
Chieko Matsumoto ◽  
Rieko Shiozawa ◽  
Shigeki Mitsunaga ◽  
Akiko Ichikawa ◽  
Rika Ishiwatari ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Detection System ◽  
Dna Detection ◽  
Pcr Detection ◽  
Acid Extraction ◽  
Nucleic Acid Extraction ◽  
Proviral Dna

scholarly journals DisCVR: Rapid viral diagnosis from high-throughput sequencing data

2019 ◽  
Vol 5 (2) ◽  
Author(s):  
Maha Maabar ◽  
Andrew J Davison ◽  
Matej Vučak ◽  
Fiona Thorburn ◽  
Pablo R Murcia ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Sequencing ◽  
Clinical Sample ◽  
High Sensitivity ◽  
Clinical Samples ◽  
Upper Respiratory Tract ◽  
Sequencing Data ◽  
High Throughput Sequencing Data ◽  
Tract Infections ◽  
Human Viruses

Abstract High-throughput sequencing (HTS) enables most pathogens in a clinical sample to be detected from a single analysis, thereby providing novel opportunities for diagnosis, surveillance, and epidemiology. However, this powerful technology is difficult to apply in diagnostic laboratories because of its computational and bioinformatic demands. We have developed DisCVR, which detects known human viruses in clinical samples by matching sample k-mers (twenty-two nucleotide sequences) to k-mers from taxonomically labeled viral genomes. DisCVR was validated using published HTS data for eighty-nine clinical samples from adults with upper respiratory tract infections. These samples had been tested for viruses metagenomically and also by real-time polymerase chain reaction assay, which is the standard diagnostic method. DisCVR detected human viruses with high sensitivity (79%) and specificity (100%), and was able to detect mixed infections. Moreover, it produced results comparable to those in a published metagenomic analysis of 177 blood samples from patients in Nigeria. DisCVR has been designed as a user-friendly tool for detecting human viruses from HTS data using computers with limited RAM and processing power, and includes a graphical user interface to help users interpret and validate the output. It is written in Java and is publicly available from http://bioinformatics.cvr.ac.uk/discvr.php.

scholarly journals Establishment of a Gene Detection System for Hotspot Mutations of Hearing Loss

2018 ◽  
Vol 2018 ◽  
pp. 1-8
Author(s):  
Chao Wang ◽  
Shengzhou Wang ◽  
Hongyan Chen ◽  
Daru Lu
Keyword(s):  
Hearing Loss ◽  
Screening Program ◽  
Detection System ◽  
High Sensitivity ◽  
Extraction Process ◽  
Clinical Samples ◽  
12S Rrna ◽  
Gene Detection ◽  
Newborn Hearing ◽  
Hotspot Mutations

Hearing loss is an etiologically heterogeneous trait with a high incidence in China. Though conventional newborn hearing screening program has been widely adopted, gene detection can significantly improve the means of early discovering genetic risk factors. Thus, simple and efficient methods with higher sensitivity and lower cost for detecting hotspot mutations of hearing loss are urgently requested. Here we established a mutation detection system based on multiple fluorescent probe technique, which can detect and genotype nine hotspot mutations of four prominent hearing loss-related genes in two reactions on a four-channel real-time PCR instrument, includingGJB2(rs750188782, rs80338943, rs1110333204, and rs80338939),GJB3(rs74315319),SLC26A4(rs111033313 and rs121908362), andmtDNA 12S rRNA(rs267606617 and rs267606619). This system is with high sensitivity that enables detecting as low as 10 DNA copies samples per reaction. A comparison study in 268 clinical samples showed that the detection system had 100% concordance to Sanger sequencing. Besides, blood and saliva samples can be directly detected without DNA extraction process, which greatly simplifies the manipulation. The new system with high sensitivity, accuracy, and specimen type compatibility can be expectedly a reliable tool in clinical application.

scholarly journals Rapid ultrasensitive diagnosis of pneumonia caused by Acinetobacter baumannii using a combination of enrichment and phage-based qPCR assay

2020 ◽  
Author(s):  
Jun Luo ◽  
Mengwei Jiang ◽  
Jin Xiong ◽  
Junhua Li ◽  
Hongping Wei ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Enrichment Culture ◽  
Detection System ◽  
Sputum Sample ◽  
High Sensitivity ◽  
Rapid Identification ◽  
Clinical Method ◽  
Real Time Quantitative Pcr ◽  
Live Bacteria ◽  
Infection Patient

Abstract Background Accurate and rapid identification of ventilator associated or nosocomial pneumonia caused by Acinetobacter baumannii ( A. baumannii ) could improve the treatment.Methods In current study, we developed a phage-based real-time quantitative PCR (qPCR) combined with enrichment culture for rapid and specific detection of viable A. baumannii in sputum from lung infections. Through short-term plate incubation, bacteria can be enriched and the DNA polymerase reaction disturbance of sputum can be decreased greatly. This approach is based on detecting phage replication in live A. baumannii cells through Taqman qPCR.Results Through the built detection system, down to 1 CFU of A. baumannii can be detected within 6 h in spiked sputum samples without any steps of bacteria isolation and DNA extraction. The established method was then applied to detecting both A. baumannii in simulated sputum with 100% agreement with the spiked amount of the bacteria and one clinical sputum sample from an 80-year-old male lung infection patient caused by A. baumannii with perfect accuracy, demonstrating that the assay developed in this study has the merits of high rapidity, high sensitivity, good specificity and being able to detect live bacteria not dead bacteria.Conclusions The assay is a potentially clinical method for diagnosis of bacterial pneumonia infection caused by A. baumannii or other bacterial infection in sputum or complicated samples through switching to other types of phages.

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