Oropouche orthobunyavirus: Genetic characterization of full-length genomes and development of molecular methods to discriminate natural reassortments

SUMMARYThe development of simple, sensitive and rapid methods for the detection and identification ofToxoplasma gondiiis important for the diagnosis and epidemiological studies of the zoonotic disease toxoplasmosis. In the past 2 decades, molecular methods based on a variety of genetic markers have been developed, each with its advantages and limitations. The application of these methods has generated invaluable information to enhance our understanding of the epidemiology, population genetics and phylogeny ofT. gondii. However, since most studies focused solely on the detection but not genetic characterization ofT. gondii, the information obtained was limited. In this review, we discuss some widely used molecular methods and propose an integrated approach for the detection and identification ofT. gondii, in order to generate maximum information for epidemiological, population and phylogenetic studies of this key pathogen.

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Genetic characterization of dengue virus type 1 isolated in Brunei in 2005–2006

Journal of General Virology ◽

10.1099/vir.0.005306-0 ◽

2009 ◽

Vol 90 (3) ◽

pp. 678-686 ◽

Cited By ~ 6

Author(s):

Osmali Osman ◽

Mun Yik Fong ◽

Shamala Devi Sekaran

Keyword(s):

Dengue Virus ◽

Genetic Characterization ◽

The Philippines ◽

Full Length ◽

Envelope Gene ◽

Commercial Laboratory ◽

Pcr Products ◽

Primer Sets ◽

Genotype V

The full-length genomes of two DENV-1 viruses isolated during the 2005–2006 dengue incidents in Brunei were sequenced. Twenty five primer sets were designed to amplify contiguous overlapping fragments of approximately 500–600 base pairs spanning the entire sequence of the genome. The amplified PCR products were sent to a commercial laboratory for sequencing and the nucleotides and the deduced amino acids were determined. Sequence analysis of the envelope gene at the nucleotide and amino acid levels between the two isolates showed 92 and 96 % identity, respectively. Comparison of the envelope gene sequences with 68 other DENV-1 viruses of known genotypes placed the two isolates into two different genotypic groups. Isolate DS06/210505 belongs to genotype V together with some of the recent isolates from India (2003) and older isolates from Singapore (1990) and Burma (1976), while isolate DS212/110306 was clustered in genotype IV with the prototype Nauru strain (1974) and with some of the recent isolates from Indonesia (2004) and the Philippines (2002, 2001). In the full-length genome analysis at the nucleotide level, isolate DS06/210505 showed 94 % identity to the French Guyana strain (1989) in genotype V while isolate DS212/110306 had 96 % identity to the Nauru Island strain (1974) in genotype IV. This work constitutes the first complete genetic characterization of not only Brunei DENV-1 virus isolates, but also the first strain from Borneo Island. This study was the first to report the isolation of dengue virus in the country.

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