•9 1 Sab d vac-Cpn vac-Cpn vac-Cpn vac-Tia vac-Wye mpv-Zai mpv-Zai mpv-Congo mpv-77-666 mpv-79-005 mpv-Zai mpv-V96-l-16 ect-Mos cml-Som cml-Som cml-Som cml-Bsc _i cpv-Bri cpv-Bri

ivac-Cpn -j vac-Cpn vac-Cpn " vac-Wye mpv-Zai mpv-Zai mpv-Zai mpv-Congo mpv-77-666 mpv-79-005 mpv-V96-l-16

Icml-Som cml-Som cml-Som

Fig. 1 A-H. Dendrograms of the restriction fragment-length polymorphism (RFLP) profiles of camelpox Somalia (cml-Som), camelpox BSC-40 (cml-BSC), cowpox Brighton (cpv-Bri), ectromelia Moscow (ect-Mos), monkeypox 77-666 (mpv-77-666), monkeypox 79-005 (mpv-79-005), monkeypox Congo-8 (mpv-Congo), monkeypox V96-I-16 (mpv-V96-I-16), monkeypox Zaire (mpv-Zai), raccoonpox (rcn), Vaccinia Copenhagen (vac-Cpn), Vaccinia Tian-Tan (vac-Tia), and Vaccinia Wyeth (vac-Wye) viruses of segments 1 to 8. The similarity coefficient Sab values are indicated on the horizontal axes.

Figure 1 illustrates one LPCR-RRFLP approach using the LPCR primers listed in Table 1 and following the guidelines described above. The experiments were carried out with genomic DNAs from different strains of camelpox, cowpox, ectromelia, cotia, monkeypox, raccoonpox, and Vaccinia viruses. As shown, it was possible to classify every sample within each species. The SAB ranges of the different strains within a species were 0.53-1, 0.89-1, 0.78-1, 0.86-1, 0.89-1, 0.71-1, and 0.74-1 in segments 1, 2, 3, 4, 5, 6, and 7, respectively. The lowest SAB value within the same species (0.53) was observed with Vaccinia Copenhagen and Vaccinia Tian Tan in segment 1. The SAB ranges of different species were 0-0.31, 0.15-0.67, 0.21-0.7, 0.43-0.63, 0.13-0.86, 0.31-0.83, and 0.31-0.61 in segments 1, 2, 3, 4, 5, 6, and 7, respectively. Because only one species, Vaccinia Copenhagen, was amplified in segment 8, it was not possible to estimate the SAB values with other species or strains, but the SAB value for the different replicates of the same strain was 1.0. These results suggest that closely related species and strains can be grouped and identified using any of the eight segments. However, because lower SAB values were obtained with the flanking segments 1, 2, 7, and 8 than with the central segments 3, 4, 5, and 6, we concluded that genotyping with the terminal segments can be useful for strain differentiation whereas genotyping with the central segments can be useful for species differentiation.

Orthopoxvirus genus may emerge as natural, incidental, or deliberate health threats. Therefore, the development of rapid, sensitive, and specific methods for identification and classification of these viruses is critically important to prepare and deter against their potential resurgence. Among the modern methods for rapid, sensitive, and specific identification of orthopox viruses are real-time PCR, RFLP analysis, and microarray analysis. A method using LPCR-RFLP analysis is described to illustrate the utility of this approach in whole-genome typing of Orthopoxvirus. Using this approach, and other new approaches, could discriminate Orthopoxvirus species and strains, and possibly identify genetically mutated or altered viruses.

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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