Understanding Nucleic Acid Amplification Techniques in the Detection of Influenza viruses in Developing Countries
Authors: AbdulAzeez A. Anjorin*1,2,3, Olumuyiwa B. Salu2,3, Robert K. Obi2,3,4, Bamidele O. Oke2,3, Akeeb OB.Oyefolu1, Wellington A. Oyibo2, Sunday A. Omilabu2,3
Affiliations:
1. Department of Microbiology (Virology Research), Lagos State University, Ojo, Lagos, Nigeria
2. Department of Medical Microbiology and Parasitology, College of Medicine, University of Lagos, Nigeria
3. Virology Research Laboratory, Department of Medical Microbiology and Parasitology, College of Medicine/ Lagos University Teaching Hospital (LUTH), Idi-Araba, Nigeria
4. Department of Microbiology, Federal University of Technology, Owerri, Imo, Nigeria
Abstract
factor in the WHO influenza Global strategies for prevention and control.
Rapid, accurate, inexpensive and portable detection systems are needed
for influenza virus diagnosis and surveillance. Such a detection system
should easily identify all the subtypes of influenza virus. Degenerate
primers and probes designed from evolutionally conserved regions for
known influenza A viruses present the best way to identify unknown
subtypes of influenza A virus by polymerase chain reaction PCR and array
techniques. The isothermal reactions, Nucleic Acid Sequencing Based
Amplification (NASBA) and Loop-mediated isothermal Amplification
(LAMP) possess great potential for influenza A virus detection especially
in developing countries. However, multiplex real-time (rT) or quantitative
(q) polymerase chain reaction (qPCR) remains a rapid, accurate and timesaving technique used for influenza virus detection.
Aim: This manuscript explained the principles of nucleic acid amplification
techniques commonly used in developing countries.
Methods: Literature search was done in NCBI PUBMED, PUBMED
Central and Google Scholar using words and phrases including “Influenzamolecular diagnosis, NAAT”, Molecular techniques/ methods, PCR, qPCR,
NASBA, LAMP, and DNA microarray.
Results: The underlining principles and basic processes involved in the
application of nucleic acid amplification techniques for the detection and
epidemiological surveillance of influenza virus were identified and grouped
under PCR (RT-PCR and qRT-PCR) and Non-PCR (LCR,
pyrosequencing, NASBA, LAMP and DNA microarray) amplifications.
Conclusion: It is hoped that by understanding the techniques and basic
principles of Nucleic acid amplifications, less expensive, and more
convenient protocols for influenza virus detection and surveillance can be
developed
Keywords: Influenza, NAAT, Molecular, PCR, qPCR, Viral diagnosis.