Researchers at the University Malaya Medical Centre in Kuala Lumpur, Malaysia, are using quantitative polymerase chain reaction (qPCR; also known as real time PCR, RT PCR or qRT-PCR) to understand more about how the chikungunya virus replicates. This could lead to better diagnostics, as well as potential targets for therapeutics.
Mosquitoes spread the chikungunya virus (CHIKV), and symptoms of infection include fever, severe joint pain and arthritis, muscle pain, headache, nausea, tiredness and rash. There is no cure, only symptomatic treatment, and it can be misdiagnosed as dengue fever, as the symptoms are similar. The word ‘chikungunya’ is based on a word from the Kimakonde language of the Makonde people that means ‘to dry up or become contorted, describing how the joint pain affects infected people.
To understand more about the viral replication kinetics, the researchers used qPCR, which allowed them to quantify the levels of amplification of the target RNA strands from the RNA virus, and follow the process in ‘real time’ (read more about qPCR in Working in real time: A focus on qPCR).
The team started by creating positive and negative strand qPCR assays designed to detect the chikungunya virus non-structural protein nsP3. The positive strand assay was a more efficient diagnostic tool than an existing assay, detecting RNA copies up to day 9, even when anti-chikungunya virus IgM was present. They found no links between levels of arthralgia and viral load.
The negative strand assay used tagged primers to increase specificity, and has potential for the study of active viral replication kinetics, which could lead to better understanding of the disease, and even potential treatments. The research was published in Diagnostic Microbiology and Infectious Disease.
BJS Biotechnologies’ has launched xxpress, its ultra-fast thermal cycler, in the UK and Germany in 2013. While this project didn’t use xxpress’ qPCR technology for real time PCR analysis, its fast turnaround (40 cycles in less than 15 minutes), allows research planning to follow the data, and its simple-to-use interface makes it easy even for people new to PCR.
Suzanne Elvidge is a freelance science, biopharma, business and health writer with more than 20 years of experience. She has written for a range of online and print publications including FierceBiomarkers, FierceDrugDelivery, European Life Science, the Journal of Life Sciences (now the Burrill Report), In Vivo, Life Science Leader, Nature Biotechnology, New Scientist, PR Week and Start-Up. She specialises in writing on pharmaceuticals, biotechnology, healthcare, science, lifestyle and green living, but can write on any topic given enough tea and chocolate biscuits. She lives just beyond the neck end of nowhere in the Peak District with her second-hand bookseller husband and two second-hand cats.