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We performed a systematic investigation, from epidemiology, pathogen isolation, virus characterization, disease reproduction by infection with the isolated virus (fulfilling Koch's postulates), to virus genome sequencing and found that the duck egg drop disease was caused by a new flavivirus, BYD virus, that is closely related to Tembusu virus.
The nomenclature system used here to represent H9N2 virus lineages includes (in order) their place of isolation, virus number, and isolation time.
Virus isolation, virus load measurement and serology were performed using these specimens.
Diagnosis of sheeppox is usually based on highly characteristic clinical signs [ 9], virus isolation, virus neutralisation test (VNT) [ 10], serological assays (e.g. ELISA) [ 11] and PCR assays [ 12].
*Virus was detected in serum samples by PCR and virus isolation; virus was detected in organs by PCR only.−, negative; +, positive.
On the basis of pathologic examinations, virus isolation, virus identification and genetic characterization, we found an association with the new DTMUV.
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As has been observed in virus isolation, viruses may be able to survive longer in differentiated epidermal tissue such as contour feathers.
Virus isolation and virus stock preparation from all the five patients studied were performed using two donor's PBMCs that were pooled.
Virus isolation rates and virus titers in the lower respiratory tract were comparable between the two viruses (P>0.05), with 75% of the 48 samples being positive for Sw/B/99.
The identification and isolation of virus strains was performed according standard procedures.
The isolation of virus by the method described here requires capture using CD44 selection.
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