|Title:||Flow Cytometry Compared with Indirect Immunofluorescence for Rapid Detection of Dengue Virus Type 1 after Amplification in Tissue Culture||Authors:||KAO, CHUAN-LIANG
CHEN, LI -KUANG
|Keywords:||Dengue;Rapid Laboratory Diagnosis;Flow Cytometry;Immunofluorescence;epidemic;Taiwan||Issue Date:||2001||Journal Volume:||v.39||Journal Issue:||n.10||Start page/Pages:||3672-7||Source:||JOURNAL OF CLINICAL MICROBIOLOGY||Abstract:||
Dengue virus (DV) was detected early in infected mosquito C 6/36 cells by using indirect immunofluorescence (IF) in conjunction with flow cytometry . Three fixation- permeabilization methods and three DV serotype 1 (DEN-1) - specific monoclonal antibodies, 8-8 (anti-E), 16-4 (anti-NS1 ), and 15F3 -1 (anti-NS1), were evaluated for the detection of DEN-1 in infected C6/36 cells. We found that these three monoclonal antibodies were capable of detecting DV in C6/36 cells as early as 24 h postinoculation by using a conventional indirect IF stain. Both 8-8 and 16-4 detected DV earlier and showed a greater number of DV-positive cells than 15F3-1. In flow cytometry, 3% paraformaldehyde plus 0.1 % Triton X-100 with 16-4, the best fixation-permeabilization method for testing DV, showed higher sensitivity (up to 1 PFU) than indirect IF stain. The higher sensitivity of 16-4 in detecting DEN-1 was found with both IF and flow cytometry . Flow cytometry, which had a sensitivity similar to that of nested reverse transcription-PCR, w than 15F3-1. In flow cytometry, 3% paraformaldehyde plus 0.1
|Appears in Collections:||醫學檢驗暨生物技術學系|
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