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Detection of HIV-1 Proviral DNA in Peripheral Blood Mononuclear Cells by Nested PCR and Biotinylated Oligonucleotide Probe Hybridization

หน่วยงาน ฐานข้อมูลวิทยานิพนธ์ไทย

รายละเอียด

ชื่อเรื่อง : Detection of HIV-1 Proviral DNA in Peripheral Blood Mononuclear Cells by Nested PCR and Biotinylated Oligonucleotide Probe Hybridization
นักวิจัย : Sudsanguan Chusacultanachai
คำค้น : -
หน่วยงาน : ฐานข้อมูลวิทยานิพนธ์ไทย
ผู้ร่วมงาน : -
ปีพิมพ์ : 2535
อ้างอิง : http://www.thaithesis.org/detail.php?id=39961
ที่มา : -
ความเชี่ยวชาญ : -
ความสัมพันธ์ : -
ขอบเขตของเนื้อหา : -
บทคัดย่อ/คำอธิบาย :

Nested-PCR technique for detection of Human Immunodeficiency virus Type 1 (HIV-1) have been developed in the present study. The three important steps

DNA extraction, amplification and detection of amplified products were investigated. DNA was extracted from peripheral blood mononuclear cells with crude lysate salting out procedure which was more advantageous than the other 3 procedures

UCLA

salting out and crude lysate especially concerning its feasibility and lity efficiency. DNA was amplified in a two-step PCR, one with outer primers and the other with inner primers nested within the outer primers. Two parameters of PCR procedure including ingradients of reaction mixture especially MaCl2 concentration, and step cycle profile had been optimized for each primer pair in order to obtain highest efficiency. The PCR product was visualized by agarose gel electrophoresis and ethidium bromide staining, then identified by dot blot hybridization using biotinylated oligonucleotide probe specific to amplified sequences. All sample were analysed with 2 sets of nested primers co mplem entary to conserved region of HIV-1 within gag and env sequences. Among 100 seropositive samples, 88 were positive after ethidium bromide staining of PCR products, 100 were positive after hybridization. Samples from 100 seronegative blood donors were absolutely negative in PCR with both detection method using isotopic and non-isotopic probes, thus demonstrated the specificity of amplification and detection. So the developed technique including nested PCR and biotinylated oligonucleotide probe hybridization will be a useful technique to detect HIV-1 proviral DNA in human blood samples especially blood donors during the window period before seroconversion and newborns of seropositive mothers which can not be determined by any available serological diagnostic assays.

บรรณานุกรม :
Sudsanguan Chusacultanachai . (2535). Detection of HIV-1 Proviral DNA in Peripheral Blood Mononuclear Cells by Nested PCR and Biotinylated Oligonucleotide Probe Hybridization.
    กรุงเทพมหานคร : ฐานข้อมูลวิทยานิพนธ์ไทย.
Sudsanguan Chusacultanachai . 2535. "Detection of HIV-1 Proviral DNA in Peripheral Blood Mononuclear Cells by Nested PCR and Biotinylated Oligonucleotide Probe Hybridization".
    กรุงเทพมหานคร : ฐานข้อมูลวิทยานิพนธ์ไทย.
Sudsanguan Chusacultanachai . "Detection of HIV-1 Proviral DNA in Peripheral Blood Mononuclear Cells by Nested PCR and Biotinylated Oligonucleotide Probe Hybridization."
    กรุงเทพมหานคร : ฐานข้อมูลวิทยานิพนธ์ไทย, 2535. Print.
Sudsanguan Chusacultanachai . Detection of HIV-1 Proviral DNA in Peripheral Blood Mononuclear Cells by Nested PCR and Biotinylated Oligonucleotide Probe Hybridization. กรุงเทพมหานคร : ฐานข้อมูลวิทยานิพนธ์ไทย; 2535.