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1
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Polymerase chain reaction (PCR)
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To detect B19V prevalence in serum samples.
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2
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Enzyme-linked immunosorbent assay (ELISA)
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To analyze anti-B19V-IgM and anti-B19V-IgG antibodies.
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3
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Direct DNA-sequencing
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To reconfirm the specificity of PCR products by sequencing.
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4
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High Pure Viral Nucleic Acid Kit (Roche, Grenzach-Wyhlen, Germany)
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To extract nucleic acid from patient sera.
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5
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GeneAmp 7300 sequence analyser (Applied Biosystems Perkin-Elmer, Foster City, CA)
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To determine B19V-DNA load in serum using quantitative real-time PCR (qPCR).
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6
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TaqMan Sequence Detection System (Applied Biosystems Perkin-Elmer, Foster City, CA)
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To perform B19V-specific quantitative real-time PCR (qPCR).
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7
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BigDye Terminator Cycle Sequencing reaction (Life Technologies Corp., USA)
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To perform sequencing reactions on purified PCR products.
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8
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B19V primers
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For PCR and sequencing to detect the B19V VP1/VP2 and NS1/VP1u regions.
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9
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Parvovirus B19-IgM ELISA kit (DxSelect FocusDiagnostics, Germany)
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To detect anti-B19V-IgM antibodies.
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10
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Parvovirus B19-IgG ELISA kit (DxSelect FocusDiagnostics, Germany)
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To detect anti-B19V-IgG antibodies.
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11
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WHO international standards for B19V-DNA (NIBSC Code 99/800)
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To determine the specificity and sensitivity of the B19V-qPCR.
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