Recombinant Parvovirus B19 Pep B Protein (aa 485-781)

Up till now, screening for human parvovirus B19 is not routine in national Egyptian blood bank strategy. Blood samples were collected from 500 healthy blood donors within the age range from 18 to 45 years old attending the blood bank of Suez Canal University Hospital, Ismailia, Egypt. Sera were separated and stored at - 20 degrees C. Serum samples were screened for anti-human parvovirus B19 IgM and IgG antibodies and B19 genome using ELISA and real-time PCR respectively. Frequency of B19 IgM and B19 IgG antibodies was 6.20%, and 80.20% respectively, and the prevalence of B19 genome was 3.00%. There is a high frequency of human parvovirus B19 among Egyptian blood donors; therefore, serological screening for B19 is warranted.

In order to establish a high-throughput identification technique that simultaneously detects six major pathogens including APP, HPS, PRRSV, Mhp, PCV-2 and CSFV, six pairs of primers and probes were designed based on the specific conservative sequences of the pathogens, a multiplex PCR system was developed, hybrid parameters were optimized, and evaluation of the technology was performed. The results showed that the present detection method had a sensitivity of 5.8 x 10(2)copies/mu L for APP, 7.8 x 10(3) copies/mu L for HPS, 6.8 x 10(3) copies/mu L for Mhp, 6.3 x 10(2) copies/mu L for PCV-2, 4.8 x 10(3) copies/mu L for PRRSV, and 5.5 x 10(2) copies/mu L for CSFV, respectively; and it produced no cross reaction against the other nine pathogens like swine-origin pseudorabies virus, porcine parvovirus, Japanese B encephalitis virus, swine vesicular disease virus, vesicular stomatitis virus, foot-and-mouth disease virus, bluetongue virus, peste des petits ruminants virus and salmonella. Application of the multiplex oligonucleotide microarray established here to testing 285 clinical blood samples indicated a single infection rate of 18.2 % (52/285) and a mixed infection rate of 6.3 % (18/285) which were consistent with the results of the sequencing verification. This technique might serve as a rapid and high-throughput method of detection for epidemic investigation and clinical diagnosis of multiple pathogens.