Application of polymerase chain reaction (PCR) in virus diagnosis and bone marrow transplantation

Abstract

Polymerase chain reaction (PCR) is a very potent method enabling the amplification of target DNA sequences at least 10(5)-10(6) fold. The efficacy of PCR is measured by its specificity, efficiency (yield quantity) and fidelity. On the basis of PCR specificity, it is possible to differentiate host genom from virus or bacterial genom if they are in the host bloodstream. It is possible to make a precise diagnosis of an infectious disease. The result of bone marrow transplantation can also be more quickly predicted than by other techniques. Prognosis of engraftment following an allogeneic bone marrow transplantation was obtained by PCR amplification of the hypervariable region (HVR) lying on the 3end of the apolipoprotein B gene. This HVR polymorphism of the largest human gene (apoB) is very informative, considering that 16 alleles are registered in the Belgrade population. It is necessary to detect the HVR genotype of donor and host before and after allogenic bone marrow transplantation. The aim is to monitor engraftment in peripheral blood samples in early stages (two or three weeks) after bone marrow transplantation. A test of detection of hepatitis C virus in hepatic patients (positive control) and in some immunoglobuline preparations, was also carried out. Because HCV is an RNA virus, a nested (two stages) PCR amplification of highly conserved 5-noncoding region was performed after a reverse transcription. Polymerase chain reaction could be used for the rapid detection of pathogens, especially those for which detection, using other methods, is difficult or unreliable. The application of PCR in diagnosis has an unprecedented success rate. By now, because of its simplicity and extreme sensitivity and also its specificity, the PCR technology is better than its competition.