Journal: Scientific reports

Article Title: Development of a diagnostic assay by three-tube multiplex real-time PCR for simultaneous detection of nine microorganisms causing acute respiratory infections.

doi: 10.1038/s41598-022-15543-6

Figure Lengend Snippet: Figure 4. Diagnostic performance evaluation of the multiplex real-time RT-PCR. A, schematic representation of flow chart of the multiplex real-time PCR assay for detection of clinical specimens. After the patient’s nasopharyngeal/throat swab is taken, the nucleic acid extracted from the sample is amplified simultaneously in three tubes by the multiplex PCR assay. If the fluorescence channel of RSV in tube A shows an amplification curve and GAPDH is detected in all three tubes, a valid sample collection can be diagnostic for RSV infection in the sample. B, the number of each pathogen detected by three-tube nine-plex real-time PCR. The total number of pathogens detected in each type of infection were as follows: Single, 7; Dual, 7; Triple, 7; Quadruple, 4. LP: Legionella pneumophila, HI: Haemophilus influenzae, MP: Mycoplasma pneumoniae, CP: Chlamydia pneumoniae, SP: Streptococcus pneumoniae.

Article Snippet: Number Species Subtypes Strain ID Supplier Description 1 Influenza A virus H1N1 ATCC VR95 American Type Culture Collection 2 H3N2 GDV124 The State Key Laboratory of Virology, Wuhan University, CHINA3 H1N1 2009 GDV108 4 H5N1 The National Institute for Food and Drug Control, CHINA Supplied as virus cultures5 H7N9 6 Influenza B virus IVB Yamataga GDV105 The State Key Laboratory of Virology, Wuhan University, CHINA7 IVB Victoria GDV104 8 Respiratory syncytial virus RSV A ATCC VR1540 American Type Culture Collection 9 RSV B ATCC VR 955 10 Adenovirus Human adenovirus 1 ATCC VR-1 American Type Culture Collection 11 Human adenovirus 2 ATCC VR-846 12 Human adenovirus 3 ATCC VR-3 13 Human adenovirus 4 ATCC VR-1572 14 Human adenovirus 5 ATCC VR-5 15 Human adenovirus 46 ATCC VR-1308 16 Human adenovirus 48 ATCC VR-1406 17 Human mastadenovirus 7 ATCC VR-7 18 Human adenovirus 55 N/A The State Key Laboratory of Virology, Wuhan University, CHINA Supplied as clinical isolates 19 Chlamydia pneumoniae ATCC VR2282 American Type Culture Collection 20 Mycoplasma pneumoniae Mycoplasma pneumoniae Somerson et al ATCC 15531 21 Streptococcus pneumoniae Streptococcus pneumoniae Serotype 19F ATCC 49619 22 Streptococcus pneumoniae (Klein) Chester ATCC 700669 23 Legionella pneumoniae Legionella pneumophila subsp. fraseri Brenner et al ATCC 33156 24 Legionella pneumophila subsp. pneumophila Brenner et al ATCC 43290 25 Haemophilus influenzae Haemophilus influenzae (Lehmann and Neumann) Winslow et al ATCC 10211 12 Vol:. (1234567890) Scientific Reports | (2022) 12:13306 | https://doi.org/10.1038/s41598-022-15543-6 real-time RT-PCR assay followed the pattern of selecting conserved gene sequence regions.

Techniques: Diagnostic Assay, Multiplex Assay, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Amplification, Fluorescence, Infection