成人社区获得性肺炎的病毒病原学及临床特征研究-study on viral etiology and clinical characteristics of adult community acquired pneumonia.docxVIP

广州医学院硕士学位论文 广州医学院硕士学位论文 Abstract Abstract - - PAGE 10 - - - PAGE 7 - Abstract Part One Establishment of a rapid cell culture system for dignosis of common respiratory viruses Background:There are several diagnostic techniques available for viral detection, including viral isolation, viral nucleic acid detection, viral antigen dectection and serum specific antibody. All these methods are routinely used for viral detection in some developed countries, but not available in majority of hospitals in mainland China. In limited hospitals that viral detection is available for clinical practice, the most commonly used methods are immunofluorescence, polymerase chain reaction (PCR), or virus specific IgM antibody in acute phase serum. Viral isolation, as the “gold standard” of diagnosis of viral infection, is rarely available for clinical application. Establishment of multiple modalities system, including molecular diagnosis, cell culture for viral isolation and convalescent serum antibodies, should be important for the clinical monitoring and diagnosis of viral respiratory infections. Objective: To establish a rapid cell culture system, which combined enhanced cell culture and direct IF, for detection of common respiratory viruses, and to test the sensitivity of this system. Methods: Nasopharyngeal swab or/and throat swab samples were collected from adults that admitted to Emergency Department of Guangdong Provincial Hospital of Traditional Chinese Medicine (Guangzhou, China) and from children that admitted to Shenzhen Children’s Hospital (Shenzhen, China) with a diagnosis of acute respiratory infection. Samples were inoculated onto MDCK, LLC-MK2 and HEp-2 cells, followed by centrifugation at 3000rpm for 60minutes, and then equal volume of cell specific medium was added. Plates were incubated at 37℃ in 5% CO2 and inspected three to four times weekly for the presence of a cytopathic effect (CPE). Hemagglutination test (HAT) was performed at 36 to 48 hours