Rapid detection of common viruses using multi-analyte suspension arrays

Abstract

A method that uses specific oligonucleotide probes coupled to a specific array of fluorescent microspheres in multi-analyte suspension arrays was employed for the detection of common viruses, such as Herpes virus (HSV), Human papillomavirus (HPV) and Hepatitis B virus (HBV). Sixteen species-specific probes and 9 sets of specific primers were designed based on conserved sequences of these viruses in the GenBank database. Serial symmetric PCR, asymmetric PCR and multiple PCR assays were employed to evaluate the sensitivity, specificity and reproducibility of multi-analyte suspension arrays analyzed on a Luminex-100 analyzer instrument. The symmetric PCR amplification of four types of HSV, four types of HPV and HBV genotypes of B, C and D, combined with their corresponding species-specific probes and specificities were completely concordant with the results from a comparative sequence analyses. There was no significant difference in the median fluorescence intensity (MFI) value between symmetric PCR and asymmetric PCR when the viral DNA concentration was above 104copies/test. Both PCR products were negative in the multi-analyte suspension arrays with viral DNA concentrations less than 103copies/test. A multi-analyte suspension array is a flexible, high-throughput, relatively simple method for rapid identification of common viruses in the clinical laboratory.