Chun Kang

Avian Influenza (H5N1) Viruses Isolated from Humans in Asia in 2004 Exhibit Increased Virulence in Mammals

ABSTRACT The spread of highly pathogenic avian influenza H5N1 viruses across Asia in 2003 and 2004 devastated domestic poultry populations and resulted in the largest and most lethal H5N1 virus outbreak in humans to date. To better understand the potential of H5N1 viruses isolated during this epizootic event to cause disease in mammals, we used the mouse and ferret models to evaluate the relative virulence of selected 2003 and 2004 H5N1 viruses representing multiple genetic and geographical groups and compared them to earlier H5N1 strains isolated from humans. Four of five human isolates tested were highly lethal for both mice and ferrets and exhibited a substantially greater level of virulence in ferrets than other H5N1 viruses isolated from humans since 1997. One human isolate and all four avian isolates tested were found to be of low virulence in either animal. The highly virulent viruses replicated to high titers in the mouse and ferret respiratory tracts and spread to multiple organs, including the brain. Rapid disease progression and high lethality rates in ferrets distinguished the highly virulent 2004 H5N1 viruses from the 1997 H5N1 viruses. A pair of viruses isolated from the same patient differed by eight amino acids, including a Lys/Glu disparity at 627 of PB2, previously identified as an H5N1 virulence factor in mice. The virus possessing Glu at 627 of PB2 exhibited only a modest decrease in virulence in mice and was highly virulent in ferrets, indicating that for this virus pair, the K627E PB2 difference did not have a prevailing effect on virulence in mice or ferrets. Our results demonstrate the general equivalence of mouse and ferret models for assessment of the virulence of 2003 and 2004 H5N1 viruses. However, the apparent enhancement of virulence of these viruses in humans in 2004 was better reflected in the ferret.

Outbreak of measles in the Republic of Korea, 2007: importance of nosocomial transmission.

BACKGROUND From 2002 through 2006, Republic of Korea conducted extensive measles elimination activities and declared elimination in 2006. An outbreak of measles involving 180 confirmed cases occurred during 2007. METHODS An outbreak investigation was performed and enhanced surveillance was implemented. Detailed case investigations and laboratory testing included serologic and molecular diagnostic methods. Cases were classified according to World Health Organization and national guidelines. RESULTS During 2007, 451 suspected cases were reported and 180 (40%) cases were confirmed as measles during epidemiologic weeks 14-42. Incidence during the outbreak was 3.7 cases per million persons, excluding imported cases. Most confirmed cases were reported from Seoul; 137 (76%) cases were among children <24 months old, 124 (69%) case patients had no history of measles vaccination, and 81 (45%) case patients resulted from nosocomial transmission in 6 hospitals. Community members, patients, and health care workers all contributed to measles virus transmission. Limited outbreak control measures were implemented; high population immunity likely accounted for the self-limited transmission during this outbreak. CONCLUSIONS Limited outbreaks of measles, in which nosocomial transmission can play an important role, may occur after countries have declared elimination. Timely and opportunistic vaccination may help prevent such outbreaks; high-quality surveillance is critical for their detection.

Amantadine-resistant influenza A viruses isolated in South Korea from 2003 to 2009

To investigate the frequency of amantadine resistance among influenza A viruses isolated in Korea during the 2003-2009 seasons, 369 (16.8%) 2199 A/H1N1 viruses and 780 (14.8%) of 5263 A/H3N2 viruses were randomly selected. The M2 and HA1 genes of each isolate were amplified by reverse transcription-polymerase chain reaction and followed by nucleotide sequencing. The results showed that the resistance rate to amantadine among A/H1N1 viruses increased significantly from 2004-2005 (33.3%) to 2007-2008 (97.8%) and then decreased dramatically in 2008-2009 (1.9%). The A/H1N1 isolates recently detected in 2008-2009 turned amantadine-sensitive containing two new substitutions at specific sites (S141N, G185A) in HA1. Compared with A/H1N1 viruses, the amantadine resistance among the A/H3N2 viruses increased from 2003-2004 (9.7%) to 2005-2006 (96.7%) and decreased in 2006-2007 (57.4%). During 2006-2007, both of amantadine-resistant and -sensitive A/H3N2 viruses co-circulated but clustered in different branches phylogenetically. All of A/H3N2 isolates tested during 2007-2009 appeared to cluster in the same group being resistant to amantadine.

High isolation rate of adenovirus serotype 7 from South Korean military recruits with mild acute respiratory disease

Adenovirus is a major cause of acute respiratory disease (ARD) in military recruits. When South Korean military recruits with ARD were surveyed, adenovirus was identified in 122 (61.0%) of the 200 recruits studied. Moreover, all cases of ARD involving adenovirus were caused by serotype 7.

Development of real-time PCR assays for detection and quantification of human bocavirus.

Abstract Background Human bocavirus (HBoV) is a parvovirus that has been recently detected in patients with respiratory illness. Objectives We developed a sensitive, specific, and quantitative real-time PCR assay based on the TaqMan method for HBoV detection and quantification in respiratory specimens. Study design Three individual real-time PCR assays were designed to amplify HBoV NS1, NP-1, and VP1 genes. For clinical evaluation, 506 nasal aspirates obtained from patients with acute respiratory tract infections during December 2006 to May 2007 were tested. Results Each assay had a broad dynamic range (50×107 to 5×107 copies of plasmid DNA) and high inter- and intra-assay reproducibility. The detection limit of each assay was 10 genome copies per reaction, and no crossreactivity with other major respiratory viruses or bacteria was detected. Clinical evaluation revealed that 11 (2.1%) of 506 patients diagnosed with upper respiratory tract infections, pneumonia, bronchitis, pharyngitis, or sinusitis had HBoV detected by all three assays, with viral loads ranging from 8.2×104 to 8.1×109 copies/ml of specimen. Conclusions The three assays for HBoV diagnosis and quantification are highly sensitive, specific real-time tools for the reliable epidemiological and pathogenetic study of HBoV infection.

Oseltamivir-Resistant Pandemic (H1N1) 2009 Virus, South Korea

To identify oseltamivir resistance, we analyzed neuraminidase H275Y mutations in samples from 10 patients infected with pandemic (H1N1) 2009 virus in South Korea who had influenza that was refractory to antiviral treatment with this drug. A neuraminidase I117M mutation that might influence oseltamivir susceptibility was detected in sequential specimens from 1 patient.

Influenza surveillance in Korea: establishment and first results of an epidemiological and virological surveillance scheme

Surveillance is an important component of influenza control. This report describes the establishment and first results of the Korean Influenza Surveillance Scheme (KISS), an integrated clinical and laboratory surveillance network involving 622 public health centres (PHCs) and private clinics. Sentinel physicians reported cases of influenza-like illness (ILI) weekly and forwarded specimens for virus isolation and characterization. Influenza activity during the opening 2000-2001 season was milder and delayed compared with previous years. The ILI consultation rate corresponded well with the number of influenza virus isolates, both peaking in week 10 of 2001. Influenza A(H3N2) was the dominant isolate. The peak ILI consultation rate was higher in private clinics than in PHCs (5.04 vs 1.79 cases/1000 visits). An evaluation questionnaire generated potential enhancements to the scheme. KISS appears to represent the pattern of influenza activity accurately and will have a valuable role in monitoring and preventing epidemics in Korea.

Silencing E1A mRNA by RNA interference inhibits adenovirus replication

SummaryThe adenovirus family contains 51 human serotypes, and most human adenoviruses cause widespread respiratory tract infections. Adenovirus infections can result in severe complications in some cases, such as in adenovirus type 11 infection in immunocompromised patients. However, effective treatment methods for adenovirus infections are currently unavailable. This prompted the search for antiviral agents effective against adenovirus infections. In the present study, adenovirus E1A was targeted by RNA interference (RNAi) using synthetic small interfering RNAs (siRNAs) in an attempt to inhibit viral replication, since adenovirus E1A proteins are known to be involved in the transcriptional activation of the viral and cellular genes necessary for controlling the cell cycle and viral replication. The results indicated that the siRNAs effectively reduced the amount of adenovirus E1A mRNA and the levels of replicative intermediates. Additionally, siRNA-mediated gene silencing inhibited adenovirus replication by suppressing the E1A mRNA. These results suggest that the RNAi-mediated targeting of adenovirus E1A may have a potentially therapeutic effect in controlling adenovirus infections.

Replication and pathogenesis of the pandemic (H1N1) 2009 influenza virus in mammalian models

This study aimed to characterize the replication and pathogenic properties of a Korean pandemic (H1N1) 2009 influenza virus isolate in ferrets and mice. Ferrets infected with A/Korea/01/2009 (H1N1) virus showed mild clinical signs. The virus replicated well in lungs and slightly in brains with no replication in any other organs. Severe bronchopneumonia and thickening of alveolar walls were detected in the lungs. Viral antigens were detected in the bronchiolar epithelial cells, in peribronchial glands with severe peribronchitis and in cells present in the alveoli. A/Korea/01/2009 (H1N1) virus-infected mice showed weight loss and pathological lung lesions including perivascular cuffing, interstitial pneumonia and alveolitis. The virus replicated highly in the lungs and slightly in the nasal tissues. Viral antigens were detected in bronchiolar epithelial cells, pneumocytes and interstitial macrophages. However, seasonal H1N1 influenza virus did not replicate in the lungs of ferrets, and viral antigens were not detected. Thus, this Korean pandemic (H1N1) 2009 isolate infected the lungs of ferrets and mice successfully and caused more pathological lesions than did the seasonal influenza virus.

Epidemiologic Study of Human Influenza Virus Infection in South Korea from 1999 to 2007: Origin and Evolution of A/Fujian/411/2002-Like Strains

ABSTRACT Influenza epidemics arise through the accumulation of viral genetic changes, culminating in a novel antigenic type that is able to escape host immunity. Following an outbreak of the A/Fujian/411/2002-like strains in Asia, including China, Japan, and South Korea, in 2002, Australia and New Zealand experienced substantial outbreaks of the same strains in 2003, and subsequently worldwide outbreaks occurred in the 2003-2004 season. The emergence of A/Fujian/411/2002-like strains coincided with a higher level of influenza-like illness in South Korea than what is seen at the peak of a normal season, and there was at least a years difference between South Korea and the United States. Genetic evolution of human influenza A/H3N2 viruses was monitored by sequence analysis of hemagglutinin (HA) genes collected in Asia, including 269 (164 new) HA genes isolated in South Korea from 1999 to 2007. The Fujian-like influenza strains were disseminated with rapid sequence variation across the antigenic sites of the HA1 domain, which sharply distinguished between the A/Moscow/10/1999-like and A/Fujian/411/2002-like strains. This fast variation, equivalent to approximately 10 amino acid changes within a year, occurred in Asia and would be the main cause of the disappearance of the reassortants, although the reassortant and nonreassortant Fujian-like strains circulated simultaneously in Asia.