AI Article Synopsis
- The study aimed to develop a quick and precise real-time PCR test for the early detection of human respiratory syncytial virus (hRSV) in infants and young children using specialized primers and a TaqMan probe.
- The new assay demonstrated a high sensitivity (1 x 10² cDNA copies/microl) that was 10 times more sensitive than standard PCR and showed specific detection of hRSV compared to other viruses like polio and influenza.
- Results indicated that this assay could effectively identify hRSV in about 44% of tested samples, suggesting its potential for early diagnosis and monitoring of treatment effectiveness, though it had low correlation with results from traditional ELISA methods.
Article Abstract
Objective: To Establish a rapid and objective quantitative fluorogenic real-time PCR assay for early detection of human respiratory syncytial virus (hRSV).
Methods: Two pairs of primers and one TaqMan Fluorogenic probe that are specific for the recognition of the most conservative N gene of hRSV for virus detection with LighCycler PCR in 93 nasopharyngeal secretion specimens collected from infants and young children. The assay was compared with virus isolation, routine PCR, nested PCR, and enzyme-linked immunosorbent assay (ELISA).
Results: This TaqMan assay had a sensitivity of 1 x 10(2) cDNA copies/microl with a dynamic range between 1 x 10(2) and 1 x 10(7) cDNA copies/microl, which was the same as that of nested PCR, but 10 times more sensitive than routine PCR. The specificity of the assay was evaluated by comparing hRSV with polivirus type 1, coxsackie virus type 2, influenza A, influenza B and adenovirus type 7. A PCR product of the expected size (195 bp) was produced and fluorescence signal detected for hRSV, but not for any of the other viruses. The results in LightCycler and Rotor-Gene instrument were consistent. Forty-four specimens (43.9%) were hRSV-positive with this assay and 4 (4/93,4.3%) were hRSV-positive with ELISA, showing rather low correlation between the two methods. No visible relation was found between the concentration of hRSV RNA and severity of the disease.
Conclusion: This assay is rapid, sensitive, specific and quantitative, and has the potential of wide application for early diagnosis of hRSV infection and evaluation of the therapeutic effect.
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