Multiplexed Detection Of Respiratory Pathogens Using Pcr-Massarray

Respiratory Tract Infections, caused mainly by several respiratory virus, a few bacteria, mycoplasma, Chlamydia and rickettsia and so on, are the common and frequently-occurring disease around the world. And the usual viruses are flu virus (all kinds of three types), rhinovirus, adenovirus, coronavirus and syncytial virus and so on. Their clinical manifestations are similar, such as nasal discharge and having a fever all over the body.The purpose of this research was to discuss the clinical application of the mass spectra for detecting respirovirus, establish the method of detecting many kinds of zoonosis respirovirus at the same time, and offering a specific and high-throughput method of diagnosing respirovirus infection. According the record of whole genome sequences of respirovirus in GenBank, PCR primers and extension primers were designed using bioinformatics method. The specific steps of this research is described as follow. The method of detecting respirovirus via matrix-assisted laxer desorption ionization time-of -flight mass spectrometry was established. A part of unrelated viral from the same syndrome was detected to ensure the specificity of this method. Artificially plasmids were used to ensure the sensitivity of this method.The PCR-MassArray technology was certificated by 110 nasopharyngeal swab (NPS) that infected Influenza Virus A collected from Oct.17,2009 to Nov.27,2010. The results demonstrated that the concordance rate of the PCR-MassArray technology and the real-time fluorescent quantitation technology that was recommended by WHO was 94.54% (P>0.05), the sensitivity and the specificity were 96.43% and 92.59% respectively. Besides the Influenza Virus A, thirteen other multiplicity of infection and 7 species of pathogen was checked out from these samples. The present research demonstrated that the PCR-MassArray technology was characterized significantly by high sensitivity, high specificity and high flux, which was suitable for detecting respiratory virus, especially for screening the large scale respiratory virus and the epidemiological investigation.