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1
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cerebrospinal fluid (CSF) specimens from 12 patients
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To analyze for presence of infectious agents associated with neurologic disease.
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21801629
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2
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Total nucleic acid extraction
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To obtain nucleic acids from the CSF for further random amplification.
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3
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6-nt barcodes
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Added to the 5′ end of primers for the amplification process to enable identification of samples.
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4
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High-throughput pyrosequencing on a GS FLX Titanium Platform
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To sequence the nucleic acids obtained from the CSF samples.
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5
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PARV4-specific PCR primers
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To screen all 12 original CSF samples for PARV4 presence.
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6
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Limiting dilution PCR primers
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To semi-quantify PARV4 viral loads in the CSF and serum samples.
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7
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Reed-Muench formula
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To calculate endpoint titers of 50% positivity of PCR results.
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8
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Overlapping sets of amplicons
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To span the complete genome of PARV4 for genetic characterization.
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9
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Serologic assay for PARV4 antibodies
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To screen serum for immunoglobulin (Ig) G and IgM PARV4-specific antibodies.
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