Abi verity 96 thermal cycler

Total nucleic acids (DNA/RNA) were extracted from approximately 1 mL CSF specimens using the nucleic acid extraction kit (Ningbo Health Gene Technology, Ningbo, China) on the automated extraction workstation Smart LabAssist-16/32 (Taiwan Advanced Nanotech Inc, Taiwan), according to the manufacturer's instructions. The nucleic acid was subjected to multiplex amplification for all specimens using Meningitis/Encephalitis Pathogen Multiplex Detection Kit (Ningbo Health Gene Technology, Ningbo, China) on ABI Verity 96 Thermal Cycler (Thermo Fisher Scientific, Carlsbad, CA, USA). The PCR product was subjected to capillary electrophoresis and fragment analysis using a 3500Dx Genetic Analyzer (Thermo Fisher Scientific, Carlsbad, CA, USA) according to the manufacturer's protocol (Supplementary Figure S1). The procedure details had been reported previously (27 (link)). The panel was designed to detect 18 pathogens within 4 h.

The nucleic acids of each sample were subjected to multiplex amplification using a Respiratory Pathogen Multiplex Detection Kit (Ningbo Health Gene Technology, Ningbo, China) on an ABI Verity 96 Thermal Cycler (Thermo Fisher Scientific, Carlsbad, CA, USA). The PCR product was then subjected to capillary electrophoresis and fragment analysis using a 3500Dx Genetic Analyzer (Thermo Fisher Scientific, Carlsbad, CA, USA) according to the manufacturer’s protocol. The panel was specifically designed to detect 12 pathogens, namely, influenza A (H1N1) 2009 virus (09H1N1), seasonal H3N2 virus (H3N2), influenza B virus (InfB), human adenovirus (HADV), HBoV, human coronavirus (HCOV; 229E, OC43, NL63, HKU1), human metapneumovirus (HMPV), HPIV, HRV, human respiratory syncytial virus (HRSV), mycoplasma pneumonia (Mp) and chlamydia (Ch), within 4 h.