Genetic relationships determined by a DNA heteroduplex mobility assay: analysis of HIV-1 env genes

Delwart, Eric; Shpaer, Eugene G.; Louwagie, Joost; McCutchan, Francine E.; Grez, Manuel; Rübsamen‐Waigmann, Helga; Mullins, James I.

doi:10.1126/science.8235655

Cited by 741 publications

(559 citation statements)

References 37 publications

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“…To amplify the internal fragments, in a total final volume of 100µl, the same protocol was used with 2µl of first round products as template. The reference subtypes (obtained from WHO) were amplified using internal primers (second round) and 10ng of plasmid as template 5 .…”

Section: Patientsmentioning

confidence: 99%

Molecular epidemiology of HIV-1 in Rio Grande, RS, Brazil

Martínez

Barbosa

Ferreira

et al. 2002

Rev. Soc. Bras. Med. Trop.

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We conducted a molecular epidemiological study to investigate HIV-1 strains in Rio Grande, southern Brazil, searching for an association with transmission mode and risk behavior. Patients (185) identified at an AIDS treatment reference Hospital, from 1994 to 1997, were included; from which 107 blood samples were obtained. Nested PCR was realized once for each sample; for amplified samples (69) HIV subtypes were classified using the heteroduplex mobility assay. Subtypes identified were B (75%), C (22%) and F (3%). All infections with C were diagnosed after 1994. Comparing patients with B and C, no differences were detected regarding demographic, clinical and laboratory characteristics; survival analysis did not reveal differences in HIV to AIDS evolution. A higher proportion of injecting drug users, IDU (not significant, p<.07) was found among those with C. This suggests that C may have been introduced in this area through IDU, and is being spread, probably by their sexual partners, to persons with other risk practices.

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Section: Patientsmentioning

confidence: 99%

Molecular epidemiology of HIV-1 in Rio Grande, RS, Brazil

Martínez

Barbosa

Ferreira

et al. 2002

Rev. Soc. Bras. Med. Trop.

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“…The mixed PCR products were denatured at 94 0 C for two minutes in a Tec gene thermocycler (Care Biomedicals) and reannealed quickly on wet ice. Samples were mixed with gel loading dye prior to loading on the gel [9]. Electrophoresis was done using 5% non-denaturing acrylamide gel (using a 30% acrylamide, 0.8% bisacrylamide stock) in TBE buffer, running at 250 volts for three hours and …”

Section: Methodsmentioning

confidence: 99%

“…The heteroduplex mobility ratio was calculated from the mobility distance of the heteroduplexes and homoduplexes from the well bottom [9].…”

Section: Methodsmentioning

confidence: 99%

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Identification of Human Immunodeficiency Virus Type-1 Subtypes by Heteroduplex Mobility Assay

Sahni

Gupta

Nagendra

et al. 2007

Medical Journal Armed Forces India

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Background: Human immunodeficiency virus type-1 (HIV-1) has developed marked genomic sequence differences over the course of an epidemic because of an error prone reverse transcriptase (RT), which rapidly incorporates mutations resulting in genomic diversity, altered cell tropism, immune escape and variable resistance to antiretroviral drugs. The best preventive strategy for HIV control is development of an efficacious prophylactic vaccine using the most appropriate (antigenically related) subtypes. On the basis of phylogenetic analysis, HIV strains can be separated into major group "M" consisting of genetic subtypes A-K, "N", the new group and "O", the outlier group.

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“…de acrilamida e permite o estudo de um maior número de amostras, num tempo menor, do que a técnica de seqüenciamento (Delwart et al, 1993).…”

Section: Ester Cerdeira Sabinounclassified