Use of reverse transcription – polymerase chain reaction (RT-PCR) and dot blot hybridization with biotinylated labeled DNA probe techniquws to detect human immunodeficiency virus (HIV) in blood serum  

Oleh:

Rosilawati, Maria Lina ; Bela, Budiman

Jenis: Article from Journal - ilmiah nasional - terakreditasi DIKTI - non-atma jaya
Dalam koleksi: Universa Medicina vol. 26 no. 03 (Jul. 2007), page 111.
Topik: HIV; RT-PCR; dot blot hybridization; blood serum

Ketersediaan

Perpustakaan FK Nomor Panggil: U01.K Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

Lihat Detail Induk

Isi artikel

BACKGROUND Human immunodeficiency virus (HIV) in blood serum can be detected by means of molecular biology techniques such as reverse transcription – polymerase chain reaction (RT-PCR) and dot blot hibridization with a biotinylated labeled DNA probe. The techniques subsequently can be applied to screen HIV from blood of biological tissues such as radiation sterilized amnions and. allografts especially from National Nuklir Energy Agency Research Tissue Bank. Purpose of this research is, to detect HIV by comparing the sensitivity of the RT-PCR- dot blot hybridization with a biotinylated labeled DNA probe and RT-PCR-agarose gel electrophoresis methods. METHODS Samples used in this research were blood sera from Fatmawati Drug Dependent Hospital. The amount of the samples were 55 blood sera consisted of 5 negative and 50 positive HIV samples which were analysed with the rapid test and enzyme linked immunoassay (ELISA) serological assays, respectively. HIV RNA in blood serum was extracted by means of RNA viral extraction kit while one step RT-PCR technique was performed for DNA amplification process. RESULTS Results of this research revealed that the HIV positive could be detected on 43 of 55 samples either by RT-PCR agarose gel electrophoresis or RT-PCR dot blot hybridization techniques. The detection result of RT-PCR dot blot hybridization appeared much more clear than RT-PCR agarose gel electrophoresis techniques. Several samples with positive HIV appeared as thick black dots on the film whereas on agarose gel the same samples performed as thin DNA bands. CONCLUSION Based on the results obtained, the RT-PCR dot blot using biotinylated labeled probe techniques are more sensitive than RT-PCR agarose gel electrophoresis techniques to detect HIV in blood serum. Keywords : HIV, RT-PCR, dot blot hybridization, blood serum

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