Minal, Noor Hamizah
(2013)
Evaluation of primers for the detection of HIV-1 viruses from
Kelantan by using Reverse Transcription Polymerase Chain
Reaction (RT-PCR).
Project Report.
Pusat Pengajian Sains Kesihatan, Universiti Sains Malaysia.
(Submitted)
Abstract
Human immunodeficiency virus type 1 (HIV-1) has been recognized as the causative
agent of acquired immunodeficiency syndrome (AIDS). Approximately 42 million
people carrying the virus at present, but its case fatality rate is close to 100%, making it
an infection of devastating ferocity. Since the evolutionary change of the virus is at rapid rate, it imposes a challenge on HIV diagnostic development, vaccine
development, antiretroviral drug sensitivity and drug resistance. Rapid and sensitive
methods for the detection of HIV-1 viruses would be valuable in controlling this
disease. Currently, nucleic acid amplification tests such as reverse transcription
polymerase chain reaction (RT-PCR) and real-time PCR (rtPCR) are widely used in
detecting HIV-1 viruses. These tests are commonly applied in blood donation screening
and early detection of HIV-1 in infants. High sequence variations of the HIV-1 viruses
from Kelantan have led to the failure to obtain the amplification product using
published primers. Therefore, in this study, a few set of primers were designed based on
the sequence alignments of published HTV-1 viruses from Malaysia and Kelantan which
were retrieved from NCBI genebank. The primers were evaluated on 30 confirmed HIV-1 positive samples from Kelantan, Malaysia. HIV LTRF2 and HIV LTR2 were chosen as the best primers to detect these viruses since they were successfully amplified
all the 30 samples used in this study. The primers were further evaluated their
sensitivity and specificity against 3 viruses which are Japanese encephalitis (JE),
Chikungunya and Western encephalitis virus (WEE) where no expected band yielded.
These primers were sensitive and specific enough in detecting HIV-1 viruses from
Kelantan. They were further evaluated for use in the real-time PCR and can be used in
this test. Thus, these primers could be potentially use for the future diagnosis of HIV-1
viruses from Kelantan either by conventional or real time PCR.
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