Back to Search
Start Over
Real-time nucleic acid sequence-based amplification is more convenient than real-time PCR for quantification of Plasmodium falciparum
- Source :
- Journal of Clinical Microbiology, 43, 1, pp. 402-5, Journal of Clinical Microbiology, 43, 402-5, Journal of clinical microbiology, 43(1), 402-405. American Society for Microbiology
- Publication Year :
- 2005
-
Abstract
- Determination of the number of malaria parasites by routine or even expert microscopy is not always sufficiently sensitive for detailed quantitative studies on the population dynamics of Plasmodium falciparum , such as intervention or vaccine trials. To circumvent this problem, two more sensitive assays, real-time quantitative nucleic acid sequence-based amplification (QT-NASBA) and real-time quantitative PCR (QT-PCR) were compared for quantification of P. falciparum parasites. QT-NASBA was adapted to molecular beacon real-time detection technology, which enables a reduction of the time of analysis and of contamination risk while retaining the specificity and sensitivity of the original assay. Both QT-NASBA and QT-PCR have a sensitivity of 20 parasites/ml of blood, but QT-PCR requires a complicated DNA extraction procedure and the use of 500 μl of venous blood to achieve this sensitivity, compared to 50 μl of finger prick blood for real-time QT-NASBA. Both techniques show a significant correlation to microscopic parasite counts, and the quantification results of the two real-time assays are significantly correlated for in vitro as well as in vivo samples. However, in comparison to real-time QT-PCR, the results of real-time QT-NASBA can be obtained 12 h earlier, with relatively easy RNA extraction and use of finger prick blood samples. The prospective development of multiplex QT-NASBA for detection of various P. falciparum developmental stages increases the value of QT-NASBA for malaria studies. Therefore, for studies requiring sensitive and accurate detection of P. falciparum parasites in large numbers of samples, the use of real-time QT-NASBA is preferred over that of real-time QT-PCR.
- Subjects :
- Microbiology (medical)
Plasmodium falciparum
Population
Polymerase Chain Reaction
Sensitivity and Specificity
Auto-immunity, transplantation and immunotherapy [N4i 4]
Molecular beacon
parasitic diseases
Animals
Humans
Multiplex
Malaria, Falciparum
education
Self-Sustained Sequence Replication
education.field_of_study
biology
Poverty-related infectious diseases [N4i 3]
Reproducibility of Results
DNA, Protozoan
biology.organism_classification
DNA extraction
Molecular biology
Real-time polymerase chain reaction
Nucleic acid
Parasitology
RNA extraction
Microbial pathogenesis and host defense [UMCN 4.1]
Infection and autoimmunity [NCMLS 1]
Subjects
Details
- ISSN :
- 00951137
- Database :
- OpenAIRE
- Journal :
- Journal of Clinical Microbiology, 43, 1, pp. 402-5, Journal of Clinical Microbiology, 43, 402-5, Journal of clinical microbiology, 43(1), 402-405. American Society for Microbiology
- Accession number :
- edsair.doi.dedup.....6a11288c1441dea5f55d5eb099b3872c