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. 2019 Oct 10;15(9):640-645.
doi: 10.6026/97320630015640. eCollection 2019.

Characterization of influenza A(H1N1)pdm09 isolates of Peru using HRM, a post PCR molecular biology method

Priscila Lope  1   2 , Huaringa Maribel  1 , Mayta Egma  2 , Bailon Henri  3 , Padilla Carlos  3
Affiliations

Characterization of influenza A(H1N1)pdm09 isolates of Peru using HRM, a post PCR molecular biology method

Priscila Lope et al. Bioinformation. .

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.

Keywords: Genotype; H1N1 Influenza Virus Subtype A; High Resolution Melting.

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Conflict of interest statement

The authors declare no conflicts of interest

Figures

Figure 1
Figure 1
Normalized High-Resolution Melting profiles of Hemagglutinin gene. A, B, C, and D represent the profiles detected in influenza A(H1N1)pdm09 isolates of Peru during the year 2015-2016.
Figure 2
Figure 2
Normalized High-Resolution Melting profiles of Matriz gene. a, b, and c represent the profiles detected in influenza A(H1N1)pdm09 isolates of Peru during the year 2015-2016.
Figure 3
Figure 3
Distribution of genotypes of influenza A(H1N1)pdm09 virus during the years 2015-2016, using the HRM technique using the Matrix and Hemagglutinin genes. Size of circles represents the number of samples according to the scale. The map of Peru is divided by lines that represent the political division of the country. Map derived from QGIS 3.8.2-Zanzibar
See this image and copyright information in PMC

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