Rapid detection of Orthopoxvirus by semi-nested PCR directly from clinical specimens: A useful alternative for routine laboratories

摘要: Orthopoxvirus (OPV) has been associated with worldwide exanthematic outbreaks, which have resulted in serious economic losses as well impact on public health. Although the current classical and molecular methods are useful for diagnosis of OPV, they largely inaccessible to unsophisticated clinical laboratories. The major reason inaccessibility is that require both virus isolation DNA manipulation. In this report, a rapid, sensitive low-cost semi-nested PCR method described detection OPV directly from specimens. A set primers was designed amplify conserved vgf gene. most thermal chemical conditions were selected minimum non-inhibitory dilutions determined. More than 100 Brazilian Vaccinia (VACV) field specimens tested using order confirm its applicability. Cowpox also detected by ear scabs scarified Balb/c mice. addition, highly VACV murine blood excreta, among suggested reservoirs OPV. Together, these data suggest can be used initial screening routine diagnostic laboratory method.