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1
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Parvo B19 PCR Kit (Abbott Laboratories, IL, USA)
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Routine laboratory testing to detect hB19V infection.
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31401693
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2
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EasyMag platform (bioMérieux, Marcy l’étoile, France)
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DNA extraction from clinical samples.
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3
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Forward primer for first nested PCR step (5’-CACA AAG CTG GGA CCA CTA TGA A-3’)
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First step amplification in nested PCR targeting NS1 gene of hB19V.
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4
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Reverse primer for first nested PCR step (5’-GCG ACT TCC GGT AAA CTT CCT T-3’)
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First step amplification in nested PCR targeting NS1 gene of hB19V.
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5
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Forward primer for second nested PCR step (5’-TTG TAC GCT AAC TTG CCC AGG CTT-3’)
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Second step amplification in nested PCR targeting NS1 gene of hB19V.
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6
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Reverse primer for second nested PCR step (5’-CAT TTG TCG GAA GCC CAG TTT CCT-3’)
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Second step amplification in nested PCR targeting NS1 gene of hB19V.
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7
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dNTPs
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Building blocks for DNA synthesis during PCR.
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8
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Polymerase
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Enzyme to synthesize new DNA strands during PCR.
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9
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Healthcare Bio-Sciences, Uppsala, Sweden purification column
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Purification of PCR amplicons before sequencing.
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10
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Applied Biosystems 3130 Genetic Analyzer (Life Technologies, CA, USA)
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Sequencing of hB19V amplicons for phylogenetic analysis.
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11
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WHO reference samples
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Control to assess the detection of all currently circulating hB19V genotypes.
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12
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French NRC-V9 strain variants
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Control to confirm the range of PCR detection and sequencing.
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13
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Negative controls
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Control to confirm absence of cross-contamination or false amplification.
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14
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ClustalW package
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Aligning consensus nucleotide sequences for phylogenetic analysis.
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