ABSTRACT
Several novel viruses have been recently identified in respiratory samples. However, the epidemiology of these viruses in tropical countries remains unclear. The aim of the present study was to provide an overview of the epidemiology of novel respiratory viruses, including human metapneumovirus, human bocavirus, new subtypes of human coronavirus (NL63 and HKU1), KI virus, WU virus, and Melaka virus in the Philippines, a tropical country. Nasopharyngeal aspirates from 465 patients with influenza-like illness were collected in 2006 and 2007. Reverse transcription polymerase chain reaction (RT-PCR) and PCR were performed to detect viruses from culture-negative specimens. Human metapneumovirus, human bocavirus, human coronavirus HKU1, KI virus, and WU virus were detected for the first time in the Philippines; Melaka virus was not found.
Subject(s)
Nasopharynx/virology , Respiratory Tract Infections/epidemiology , Respiratory Tract Infections/virology , Virus Diseases/epidemiology , Virus Diseases/virology , Adolescent , Adult , Child , Child, Preschool , Coronavirus/isolation & purification , Female , Human bocavirus/isolation & purification , Humans , Infant , Male , Metapneumovirus/isolation & purification , Middle Aged , Philippines/epidemiology , Polymerase Chain Reaction , Polyomavirus/isolation & purification , Reoviridae/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction , Young AdultABSTRACT
Reassortment, which is the rearrangement of viral gene segments in a host cell infected with two different viruses, is an important mechanism for the evolution of influenza viruses. Mixed infections with multiple virus types could lead to reassortment. To better understand the occurrence of quasispecies in a single host, we investigated mixed infections in individual isolates of seasonal influenza A viruses using amantadine sensitivity as a marker. We cultured viruses with amantadine and performed sequencing, restriction fragment length polymorphism analysis, cloning, and quantitative PCR to detect mixed populations. Culturing with amantadine showed evidence of a high number of mixed populations, while the other assays could hardly detect mixed populations. The existence of quasispecies in each isolate was common. However, the proportion of these, which can be less than 1%, is too low to be detected by conventional methods. Such mixed populations in which reassortment occurs may have a significant role in the evolution of viruses.
Subject(s)
Influenza A Virus, H1N1 Subtype/classification , Influenza A Virus, H1N1 Subtype/genetics , Influenza A Virus, H3N2 Subtype/classification , Influenza A Virus, H3N2 Subtype/genetics , Influenza, Human/virology , Nasopharynx/virology , Reassortant Viruses/isolation & purification , Amantadine/pharmacology , Animals , Antiviral Agents/pharmacology , Cell Line , Cloning, Molecular , DNA Fingerprinting , Dogs , Humans , Influenza A Virus, H1N1 Subtype/isolation & purification , Influenza A Virus, H3N2 Subtype/isolation & purification , Molecular Sequence Data , Polymorphism, Restriction Fragment Length , RNA, Viral/genetics , Reassortant Viruses/classification , Reassortant Viruses/genetics , Sequence Analysis, DNA , Virus Cultivation/methodsABSTRACT
The frequency of the amantadine-resistant H1N1 influenza A virus has been increasing since the 2005-2006 season. It is unclear whether reassortment was involved in this trend. Here, we show that cocirculation of amantadine-resistant and -sensitive strains led to the genesis of amantadine-sensitive reassortant virus during the 2007-2008 season. Thereafter, the reassortant virus predominated. This contrasts with the trend for the H3N2 virus, in which the amantadine-resistant reassortant virus became predominant. The results suggest that it is necessary to monitor genome dynamics to understand the evolution and mechanism of the emergence and spread of antiviral resistance among influenza A viruses.
