Coupling multiplex RT-PCR to a gene chip assay for sensitive and semiquantitative detection of severe acute respiratory syndrome-coronavirus.
An early and accurate diagnostic assay for severe acute respiratory syndrome (SARS) is crucial for infection control. However, most of the diagnostic methods available today, such as real-time reverse transcriptase-polymerase chain reaction (RT-PCR), require a second detection method for confirmation because they detect a single sequence region of the SARS-coronavirus (SARS-CoV). For sensitive and accurate early diagnosis, we report a novel assay system combining multiplex RT-PCR and a diagnostic gene chip to detect multiple virus-specific genomic sequences of SARS-CoV. With 53 clinical specimens, we successfully demonstrate that this technique offers not only a high-accuracy diagnosis for early SARS infection but also a semiquantitative assay.
Juang
Jyh-Lyh
JL
Division of Molecular and Genomic Medicine, National Health Research Institutes, Taipei, Taiwan. juang@nhri.org.tw
Chen
Tsan-Chi
TC
Jiang
Shih Sheng
SS
Hsiung
Chao A
CA
Chen
Wei-Chen
WC
Chen
Guang-Wu
GW
Lin
Shiang-Ming
SM
Lin
Jih-Hui
JH
Chiu
Shu-Chun
SC
Lai
Yiu-Kay
YK
D017931:DNA Primers / Q000737:chemistry; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D018075:RNA, Complementary / Q000737:chemistry; D012367:RNA, Viral / Q000032:analysis*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis* / Q000821:virology
D017931:DNA Primers; D018075:RNA, Complementary; D012367:RNA, Viral