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

Priscila Lope; Huaringa Maribel; Mayta Egma; Bailon Henri; Padilla Carlos

doi:10.6026/97320630015640

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

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

Authors

Priscila Lope^{1

2}, Huaringa Maribel¹, Mayta Egma², Bailon Henri³, Padilla Carlos³

Affiliations

¹ Laboratorio de Referencia Nacional de Virus Respiratorio, Centro Nacional de Salud Publica, Instituto Nacional de Salud, Lima, Peru.
² Laboratorio de virologia. Universidad Nacional Mayor de San Marcos. Lima. Peru.
³ Laboratorio de Referencia Nacional de Biotecnologia y Biologia Molecular. Centro Nacional de Salud Publica. Instituto Nacional de Salud. Lima. Peru.

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.