Coupling multiplex RT-PCR to a gene chip assay for sensitive and semiquantitative detection of severe acute respiratory syndrome-coronavirus

Juang, Jyh‐Lyh; Chen, Tsan‐Chi; Jiang, Shih Sheng; Hsiung, Chao A.; Chen, Wei-Chen; Chen, Guang-Wu; Lin, Shiang-Ming; Lin, Jer-Huei; Chiu, Shu-Chun; Lai, Yiu‐Kay

doi:10.1038/labinvest.3700136

Cited by 16 publications

(14 citation statements)

References 15 publications

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“…It could be designed to simultaneously detect multiple virus targets by hybridizing to many virus-specific nucleotide probes. It has, in fact, been successfully used to develop diagnostic microarrays for hepatitis C virus (10), severe acute respiratory syndrome coronavirus (SARS-CoV) (15,18), influenza virus (16), and other viruses (4). In an effort to develop such a microarray-based method of sensitively and accurately detecting multiple serotypes of the enteroviruses in clinical samples, we combined multiplex RT-PCR (MRT-PCR) and a diagnostic microarray, a strategy we had used previously to develop a SARS diagnostic microarray (15).…”

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confidence: 99%

Combining Multiplex Reverse Transcription-PCR and a Diagnostic Microarray To Detect and Differentiate Enterovirus 71 and Coxsackievirus A16

Chen

Hsiung

et al. 2006

J Clin Microbiol

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Cluster A enteroviruses, including enterovirus 71 (EV71) and coxsackievirus A16 (CA16), are known to cause hand-foot-and-mouth disease (HFMD). Despite the close genetic relationship between these two viruses, EV71 is generally known to be a more perpetuating pathogen involved in severe clinical manifestations and deaths. While the serotyping of enteroviruses is mostly done by conventional immunological methods, many clinical isolates remain unclassifiable due to the limited number of antibodies against enterovirus surface proteins. Array-based assays are able to detect several serotypes with high accuracy. We combined an enterovirus microarray with multiplex reverse transcription-PCR to try to develop a method of sensitively and accurately detecting and differentiating EV71 and CA16. In an effort to design serotype-specific probes for detection of the virus, we first did an elaborate bioinformatic analysis of the sequence database derived from different enterovirus serotypes. We then constructed a microarray using 60-mer degenerate oligonucleotide probes covalently bound to array slides. Using this enterovirus microarray to study 144 clinical specimens from patients infected with HFMD or suspected to have HFMD, we found that it had a diagnostic accuracy of 92.0% for EV71 and 95.8% for CA16. Diagnostic accuracy for other enteroviruses (non-EV71 or -CA16) was 92.0%. All specimens were analyzed in parallel by real-time PCR and subsequently confirmed by neutralization tests. This highly sensitive array-based assay may become a useful alternative in clinical diagnostics of EV71 and CA16.

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confidence: 99%

Combining Multiplex Reverse Transcription-PCR and a Diagnostic Microarray To Detect and Differentiate Enterovirus 71 and Coxsackievirus A16

Chen

Hsiung

et al. 2006

J Clin Microbiol

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“…In any case, none of the remaining human coronaviruses can be identified on clinical symptoms alone, and coinfections with other respiratory viruses are as common as with other respiratory pathogens, making it difficult to identify which is the "leading" pathogen in multiple infections [16][17][18][19][20][21][22].…”

Section: Clinical Symptomsmentioning

confidence: 99%

The Human Coronaviruses

Schildgen

2018

Advanced Techniques in Diagnostic Microbiology

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“…There are numerous detection systems ranging from gold nanoparticles tagged with short segments of DNA to multicolour optical coding for biological assays that have been achieved by embedding different-sized quantum dots into polymeric microbeads (reviewed by Jain, 2003). Comigration electrophoregrams in combination with restriction enzyme digest have been used on chip devices for discrimination and quantitation of PCR products , and semi-quantitation of SARS-coronavirus has been described (Juang et al, 2004). Certainly many challenges lie ahead in this area, particularly to obtain a cost effective, workable, user friendly interface and end point result analysis system, but if successful its potential applications in the biological setting are far reaching.…”

Section: And Beyondmentioning

confidence: 99%

Nucleic acid amplification-based techniques for pathogen detection and identification

Monis

Giglio

2006

Infection, Genetics and Evolution

134

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Nucleic acid amplification techniques have revolutionised diagnostic and research industries. Current technologies that allow the detection of amplification in real-time are fast becoming industry standards, particularly in a diagnostic context. In this review, we describe and explore the application of numerous real-time detection chemistries and amplification techniques for pathogen detection and identification, including the polymerase chain reaction, nucleic acid sequence-based amplification, strand displacement amplification and the ligase chain reaction. The emergence of newer technologies, such as lab-on-a-chip devices and photo-cleavable linkers, is also discussed.

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Coupling multiplex RT-PCR to a gene chip assay for sensitive and semiquantitative detection of severe acute respiratory syndrome-coronavirus

Cited by 16 publications

References 15 publications

Combining Multiplex Reverse Transcription-PCR and a Diagnostic Microarray To Detect and Differentiate Enterovirus 71 and Coxsackievirus A16

Combining Multiplex Reverse Transcription-PCR and a Diagnostic Microarray To Detect and Differentiate Enterovirus 71 and Coxsackievirus A16

The Human Coronaviruses

Nucleic acid amplification-based techniques for pathogen detection and identification

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