Abstract
Influenza caused by A(H1N1)pdm09 is a public health issue with severe conditions in vulnerable populations leading to death. Therefore, it is of interest to characterize and monitor influenza A(H1N1)pdm09 genotypes using High Resolution Melting (HRM), a post PCR molecular biology method. We used HRM analysis (using RotorGene Q thermocycler) to characterize A(H1N1)pdm09 genotypes from several places of Peru. RNA was purified from nasal and pharyngeal swab samples referred to LRNVR-INS, synthesized cDNA, and then the hemagglutinin gene and matrix fragment were amplified. Thus, 287 samples positive for influenza A(H1N1)pdm09 were identified across Peru where places like Lima, Piura, and Arequipa documented highest number of cases. The HRM data was analyzed and results showed different profiles which were further grouped into four genotypes for the HA (A, B, C, D) and 3 for the M (a, b, c) genes. We also report ten genotypes (I-X) of virus using combined HA (hemagglutinin) and M gene profiles representing a national geography. The prevalent genotypes are I and II with a frequency of 35.89% (103) and 29.27% (84), respectively linking with severe acute respiratory infection.
Highlights
Influenza viruses cause a highly contagious acute respiratory disease
Influenza A virus belonging to the family Orthomyxoviridae, this respiratory pathogen causes annual epidemics and occasional pandemics [1]
Influenza A viruses tend to mutate rapidly and exchange RNA segments between same or different subtypes, generating new virus variants, this rearrangement or exchange of segments between different viruses that co-infect a cell is the main contributor to the increase in diversity and responsible for the most important pandemics [2]
Summary
Influenza viruses cause a highly contagious acute respiratory disease. Influenza A virus belonging to the family Orthomyxoviridae, this respiratory pathogen causes annual epidemics and occasional pandemics [1]. The appearance of new mutations in influenza A(H1N1)pdm09 isolates were reported, that mutations generate antigenic changes in hemagglutinin and drug-resistance alteration in neuraminidase [7,8,9,10,11,12,13]. High-resolution melting (HRM) analysis showed capable identify mutations, capable of discriminating gene variations, recognizing viral genotypes [14,15,16,17,18] which is highly consistent with others methods as qPCR and genome sequencing [7,8,9,10,11,12,13, 19,20,21,22].
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