Tyramide signal amplification of biotinylated probe in dot-blot hybridization assay for the detection of parvovirus B19 DNA in serum samples

Highly sensitive assay systems are necessary for large-scale virological sc reenings. We evaluated the use of tyramide signal amplification (TSA) for b iotinylated probe in dot-blot hybridization assay to detect B19 DNA in seru m samples. The probe was constructed by PCR and directly labeled with bioti n during amplification reaction. The sensitivity of the dot-blot hybridizat ion assay with TSA detection method was evaluated in comparison with a hybr idization assay using the direct detection of biotinylated probe by strepta vidin-biotin-alkaline phosphatase substrate. The TSA detection was able to detect 1 pg of B19 DNA and proved to be 10-50 times more sensitive than the hybridization assay with the direct detection of biotinylated probe. The a nalysis of 720 serum samples by TSA detection of biotinylated probe showed that the assay may be a valid diagnostic tool in routine testing of B19 DNA in serum samples. (C) 2000 Elsevier Science B.V. All rights reserved.