Identification of dengue sequences by genomic amplification: rapid diagnosis of dengue virus serotypes in peripheral blood

Diagnosis of Dengue by Using Reverse Transcriptase-Polymerase Chain Reaction

“…However the increasing number of reported cases of dengue fever/dengue haemorrhagic fever (DF/ DHF) in Brazil , Zagne et al 1994, Souza et al 1995, Vasconcelos et al 1995 showed the need for new approaches to diagnosis. Various protocols of polymerase chain reaction (PCR) have offered obvious advantages for the detection of dengue virus (Deubel et al 1990, Morita et al 1991, Pao et al 1992, Seah et al 1995.…”

Geographic distribution and evolution of yellow fever viruses based on direct sequencing of genomic cDNA fragments

“…The oligonucleotide used for cDNA priming on RNA from West African YF strains was complementary to 5' AUGGGGGAUGCTGCUUGG-GA 3', which corresponds to nucleotides 1237 to 1256 in the 17D strain YF virus E gene. For cDNA priming of the East and Central African strains of YF virus we used the oligonucleotide 5' TCTTGGTAGGAGTGATCA-TG 3' complementary to the E protein gene from nucleotides 1433 to 1452. cDNA was obtained by reverse transcription using 0-03 pmol of infected cell RNA, 5 pmol of YF virus-specific primer and a mixture made of RT buffer, RNase inhibitor, the four deoxynucleoside triphosphates and avian myeloblastosis virus RT, as previously described (Deubel et al, 1990(Deubel et al, , 1993. cDNAs were denatured at 95 °C for 5 min and then subjected to a 30-cycle amplification (denaturation at 95 °C for 15 s, annealing at 55 °C for 60 s and elongation at 72 °C for 60 s) by PCR using 50 pmol of the same antigenomicsense primer as used for the RT reaction, 50 pmol of a second primer of genomic sense 5' CAAGCTGCATG-GGGGGCACG 3' (nucleotides 822 to 841 in the E protein gene) and a mixture containing PCR buffer, the four deoxynucleoside triphosphates and Taq poiymerase (Deubel et al, 1990(Deubel et al, , 1993.…”

Dengue: a review of the laboratory tests a clinician must know to achieve a correct diagnosis

“…This method is rapid, sensitive, simple, and if correctly standardized, it can be used for genome detection in human clinical samples, biopsies, autopsy tissues or mosquitoes [52].…”

“…A number of RT-PCR procedures that detect and identify dengue serotypes in clinical specimens have been reported [51][52][53][54][55][56][57]. These PCR methods vary somewhat in terms of the amplified gene regions of the genome, in the ways they detect RT-PCR products, and the virus typing methods.…”

“…In addition to specific amplification and restriction enzyme analyses, other studies have demonstrated that nucleotide sequencing of gene fragments amplified by RT-PCR can be used as a fast method of genetic classification of dengue virus serotypes [52,66,67].…”