Study on the use of dried blood spot in sequencing and subtyping HIV-1 DNA genome.
- Author:
Qi ZHANG
1
;
Pin-Liang PAN
;
Hong DENG
;
Lin-Hong WANG
;
Yan JIANG
Author Information
- Publication Type:Journal Article
- MeSH: DNA, Viral; blood; HIV Infections; blood; genetics; HIV-1; genetics; Humans; Polymerase Chain Reaction; Sequence Analysis, DNA; methods
- From: Chinese Journal of Epidemiology 2006;27(9):798-802
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo use dried blood spot (DBS) in studying the sequence and subtype analysis of HIV-1 genome.
METHODS2 ml whole blood containing EDTA anticoagulant from 20 HIV-1 infected patients were collected, then 80 microl blood was used to propare DBS. QIAamp Blood Mini kit and 10% Chelex100 resin extracted DNA genome from whole blood and DBS as well as nested PCR amplified specifically HIV-1 Gp41 region from the two kinds of DNA extraction. Software MAGE 3.0 was used to study the sequence and subtype of the PCR products.
RESULTSEligible 18 paired samples were analysed to show that 16 of them belonged to C54A, 97CNGX-7F, 98CN006 subtypes. The other two samples might belong to B. CN._. RL42 and B. US. 90WEAU160.
CONCLUSIONData showed that there were parallel results between the whole blood and DBS samples including subtype analysis, position of mutation and types of amino acid sequencing. Since DBS itself facilitated the collection, transportation and storage, it could be used as a measure to collect blood sample in resource limited area and to develop molecular epidemiologic research as well as early diagnosis on infant exposed to HIV.
