Rapid oral bacteria detection based on real-time PCR for the forensic identification of saliva

Jung, Ju Yeon; Yoon, Hyun Kyu; An, Sang Hyun; Lee, Jee Won; Ahn, Eu-Ree; Kim, Yeon-Ji; Park, Hyun-Chul; Lee, Kyungmyung; Hwang, Jeong Ah; Lim, Sung-Soo

doi:10.1038/s41598-018-29264-2

Cited by 58 publications

(32 citation statements)

References 51 publications

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“…Compared with the traditional detection methods, such as immunoassay, PCR is more time-saving, sensitive and accurate. After nearly 40 years development, the PCR molecular detection technology has become the primary method for detecting disease caused by pathogenic microorganisms, and has been widely used in disease detection [2,3], clinical testing [4,5], food quarantine [6,7], forensic identification [8,9], customs inspection [10] and in various other fields [11][12][13][14]. The traditional PCR instrument is large in size and inconvenient to carry; and it has a higher demand for the experimental environment, a special biological laboratory and special laboratory technicians; and its reaction is in a larger volume, consumes more energy, and is not easy to integrate.…”

Section: Introductionmentioning

confidence: 99%

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

Jiang¹,

Jiang

et al. 2020

Preprint

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KEYWORDS 2 swine disease, microfluidic chips, PCR, multiplex and on-site detection 3 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 timeconsuming 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.Corporation for their support of chip design and fabrication and microchip based PCR detection instrument. Authors' contributionsJY designed and directed the study. JS and JY perform the experiment ,collect results and analyze data. YW and BZ draft the first vision of the manuscript. KMW, CG and DXZ collect clinical serum samples. BZ critically revised the manuscripts. All the participants carefully reviewed this vision manuscript. Ethics approval and consent to participate 11Not applicable Consent for publicationNot applicable Competing interest. Publisher's NoteSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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Section: Introductionmentioning

confidence: 99%

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

Jiang¹,

Jiang

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…Compared with the traditional detection methods, PCR is more time-saving, more sensitive and more accurate. After nearly 40 years of development, the PCR molecular detection technology has become the primary method for detecting disease genes and pathogenic microorganisms, and has been widely used in disease detection [4,5], clinical testing [6,7], food quarantine [8,9], forensic identification [10,11], customs inspection [12] and in various other fields [13][14][15][16]. The traditional PCR instrument is large in size and inconvenient to carry about; it has a higher demand for the experimental environment, a special biological laboratory and special laboratory technicians; and its reaction is in a larger volume, consumes more energy, is more likely to produce by-products, and is not easy to integrate.…”

Section: Introductionmentioning

confidence: 99%

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

Zhou

Jiang²,

et al. 2020

Preprint

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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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“…Compared with the traditional detection methods, such as immunoassay, PCR is more timesaving, sensitive and accurate. After nearly 40 years development, the PCR molecular detection technology has become the primary method for detecting diseases caused by pathogenic 4 microorganisms, and has been widely used in disease detection [2,3], clinical testing [4,5], food quarantine [6,7], forensic identification [8,9], customs inspection [10] and in various other fields [11][12][13][14]. The traditional PCR instrument is large in size and inconvenient to carry; and it has a higher demand for the experimental environment, a special biological laboratory and special laboratory technicians; and its reaction is in a larger volume, consumes more energy, and is not easy to integrate.…”

Section: Introductionmentioning

confidence: 99%

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

Jiang¹,

Jiang

et al. 2020

Preprint

View full text Add to dashboard Cite

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.

show abstract

Rapid oral bacteria detection based on real-time PCR for the forensic identification of saliva

Cited by 58 publications

References 51 publications

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

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

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

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

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