• 한국동물위생학회지
• /
• 제40권1호
• /
• pp.15-20
• /
• 2017

A two-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was designed for the rapid visual detection of the M gene of all subtypes of avian influenza virus (AIV) and the H5 gene of the H5 subtype of highly pathogenic AIV (HPAIV). The reaction carried out in two tubes in a single step at $58^{\circ}C$ for 40 min, and the assay results could be visually detected by using hydroxynaphthol blue dye. Using M or H5 gene-specific primers, the assay successfully detected all subtypes or H5 subtypes of AIVs, including the Korean representative H5N1 and H5N8 HPAIVs. The detection limit of the assay was approximately $10^{2.0}$ $EID_{50}/reaction$ for the M and H5 genes of H5N1 HPAIV, respectively, and was more sensitive than that of previously reported RT-LAMP and comparable to that of real-time RT-PCR. These results suggest that the present RT-LAMP assay, with its high specificity, sensitivity, and simplicity, will be a useful diagnostic tool for surveillance of currently circulating H5 HPAIVs and other subtypes of AIV in bird population, even in under-equipped laboratories.

• 약학회지
• /
• 제34권4호
• /
• pp.224-237
• /
• 1990

A single uniform population of specific, saturable, high affinity binding site of $[^3H]QNB$ guinuclidinyl benzilate(QNB) was identified in the rat cerebral microsomes. The Kd value(37.2 pM) for $[^3H]QNB$ calculated from the kinetically derived rate constants was in agreement with the Kd value(48.9 pM) determined by analysis of saturation isotherms at various receptor concentrations. Dimenhydrinate(DMH), histamine $H_1-blocker$, increased Kd value for $[^3H]QNB$ QNB without affecting the binding site concentrations and this effect resulted from the ability of DMH to slow $[^3H]QNB-receptor$ association. Pirenzepine inhibition curve of $[^3H]QNB$ binding was shallow(nH = 0.52) indicating the presence of two receptor subtypes with high ($M_1-site$) and low($M_2-site$) affinity for pirenzepine. Analysis of these inhibition curves yielded that 68% of the total receptor populations were of the $M_1-subtype$ and the remaining 32% of the $M_2-subtype$. Ki values for the $M_1-$ and $M_2-subtypes$ were 2.42 nM and 629.3 nM, respectively. Ki values for $H_1-blockers$ that inhibited $[^3H]QNB$ binding varied with a wide range ($0.02-2.5\;{\mu}M$). The Pseudo-Hill coefficients for inhibition of $[^3H]QNB$ binding by most of $H_1-blockers$ examined except for oxomemazine inhibition of $[^3H]QNB$ binding were close to one. The inhibition curve for oxomemazine in competition with $[^3H]QNB$ was shallow(nH = 0.74) indicating the presence of two receptor populations with different affinities for this drug. The proportion of high and low affinity was 33:67. The Ki values for oxomemazine were $0.045{\pm}0.016\;{\mu}M$ for high affinity and $1.145{\pm}0.232\;{\mu}M$ for low affinity sites. These data indicate that muscarinic receptor blocking potency of $H_1-blockers$ varies widely between different drugs and that most of $H_1-blockers$ examined are nonselective antagonist for the muscarinic receptor subtypes, whereas oxomemazine might be capable of distinguishing between subclasses of muscarinic receptor.

• Journal of Microbiology and Biotechnology
• /
• 제24권5호
• /
• pp.704-713
• /
• 2014

The highly pathogenic avian influenza A (HPAI) viruses of the H5N1 subtype infect poultry and have also been spreading to humans. Although new antiviral drugs and vaccinations can be effective, rapid detection would be more efficient to control the outbreak of infections. In this study, a phage-display library was applied to select antibody fragments for HPAI strain A/Hubei/1/2010. As a result, three clones were selected and sequenced. A hemagglutinin inhibition assay of the three scFvs revealed that none exhibited hemagglutination inhibition activity towards the H5N1 virus, yet they showed a higher binding affinity for several HPAI H5N1 strains compared with other influenza viruses. An ELISA confirmed that the HA protein was the target of the scFvs, and the results of a protein structure simulation showed that all the selected scFvs bound to the HA2 subunit of the HA protein. In conclusion, the three selected scFVs could be useful for developing a specific detection tool for the surveillance of HPAI epidemic strains.

• 약학회지
• /
• 제39권6호
• /
• pp.645-653
• /
• 1995

Dibenamine inhibited [$^{3}$H]quinuclidinyl benzilate ([$^{3}$H]QNB) binding in both concentration and incubation time-dependent manners. The $IC_{50}$/ value of dibenamine for the inhibition of the specific binding of 100 pM [$^{3}$'H]QNB following incubation of cerebral microsomes with dibenamine at 37.deg. C for 15 min was 20.mu.M. Dibenamine irreversibly decreased the binding site concentration for [$^{3}$H]QNB binding without affecting the affinity of [$^{3}$H]QNB for the muscarinic receptor. Analysis of the pirenzepine inhibition curve of [$^{3}$H]QNB binding to cerebral microsomes indicated the presence of two receptor subtypes with high(M$_{1}$ receptor, Ki=5nM) and low (M$_{2}$ receptor, Ki=160nM) affinity for pirenzepine. However, dibenamine(20.mu.M) treatment under the condition employed in these experiments caused steepening of the pirenzepine competition curve. The Ki value for pirenzepine in dibenamine treated-microsomes was approximately 120nM. suggesting a selective decrease in the number of M$_{1}$ receptor. Although dibenamine also inhibited [$^{3}$H]QNB binding to ventricular microsomes with $IC_{50}$/ value of 120.mu.M, the sensitivity for dibenamine in the ventricle was much lower than that in the cerebrum. These results indicate that dibenamine at low concentrations welectively inactivates the muscarinic M$_{1}$ receptor.

• Tuberculosis and Respiratory Diseases
• /
• 제69권3호
• /
• pp.207-211
• /
• 2010

Novel influenza A (H1N1) virus is a common pathogen of febrile respiratory infection recently. Here, we report the case of a 63-year-old male patient who presented with 3 days' ongoing cough and fever. He was diagnosed with novel influenza A (H1N1) pneumonia by real-time reverse-transcriptase-polymerase-chain-reaction (rRT-PCR). During treatment for novel influenza A (H1N1), his symptoms and radiologic findings improved initially, but multiple lung nodules developed subsequently and found on chest x-ray (on the 5th hospital day). Mycobacterium abscessus was isolated repeatedly from sputum and bronchoalveolar lavage fluid. To our knowledge, this is the first reported case of Mycobacterium abscessus lung disease in a patient with H1N1 influenza pneumonia.

• 대한임상검사과학회지
• /
• 제41권4호
• /
• pp.145-152
• /
• 2009

Novel influenza A virus, subtype H1N1 of swine-lineage, has been transmitted rapidly to many regions of the world. Rapid detection of the virus is essential to instigate appropriate patient care and public health management and for disease surveillance. The aim of this study is to determine the prevalence of novel influenza A (H1N1) virus in Korea using reverse-transcription real time polymerase chain reaction (rRT-PCR). Novel H1N1 virus was detected in a total of 8,948 nasopharyngeal samples from patients with influenza-like illness throughout Korea from August to September 2009. RNA was extracted from $300{\mu}l$of sample using an RNA extraction kit (Zymo Research, CA, USA). In the present study, Genekam kit (Genekam, Duisburg, Germany) was used to detect novel H1N1 virus. Novel H1N1 virus was found in 1,130 samples from a total of 8,948 samples (12.6%). The highest frequency was found in 10- to 19-year-olds (M: 29.3% vs. F: 16.4%), followed by 20- to 29-year-olds (M: 17.9% vs. F: 15.4%), 40- to 49-year-olds (M: 6.5% vs. F: 8.1%), 50- to 59-year-olds (M: 6.0% vs. F: 5.5%), and 30- to 39-year-olds (M: 4.6% vs. F: 3.8%). The mean positive rate was higher in men than in women (M: 14.7% vs. F: 7.4%). Novel H1N1 virus showed the lowest prevalence in patients over 60 years old. The positive rate increased daily and showed a significant high peak in mid-September 2009. In 19 provinces of Korea, Cheonan (41.1%), Busan (37.3%), Gangneung (33.3%), Jinju (32.1%), Ulsan (24.6%), Deajeon (23.7%) areas showed high frequencies and other provinces were found less than 10% of novel H1N1 virus. Since reverse-transcription real time PCR assay is rapid, accurate, and convenient, it may assist public health laboratories in detecting novel H1N1 virus. Moreover, these data could be useful for the management of patients with influenza-like illness.

• Tuberculosis and Respiratory Diseases
• /
• 제70권5호
• /
• pp.397-404
• /
• 2011

Background: To evaluate chest CT findings of pandemic influenza A/H1N1 pneumonia without co-infection. Methods: Among 56 patients diagnosed with pandemic influenza A/H1N1 pneumonia, chest CT was obtained in 22 between October 2009 and Februrary 2010. Since two patients were co-infected with bacteria, the other twenty were evaluated. Predominant parenchymal patterns were categorized into consolidation, ground glass opacity (GGO), and mixed patterns. Distribution of parenchymal abnormalities was assessed. Results: Median age was 46.5 years. The CURB-65 score, which is the scoring system for severity of community acquired pneumonia, had a median of 1. Median duration of symptoms was 3 days. All had abnormal chest x-ray findings. The median number of days after the hospital visit that Chest CT was performed was 1. The reasons for chest CT performance were radiographic findings unusual for pneumonia (n=13) and unexplained dyspnea (n=7). GGO was the most predominant pattern on CT (n=13, 65.0%). Parenchymal abnormalities were observed in both lungs in 13 cases and were more extensive in the lower lung zone than the upper. Central and peripheral distributions were identified in ten and nine cases, respectively. One showed diffuse distribution. Peribronchial wall thickening was found in 16 cases. Centrilobular branching nodules (n=7), interlobular septal thickening (n=4), atelectasis (n=1), pleural effusion (n=5), enlarged hilar and mediastinal lymph nodes (n=6 and n=7) were also noted. Conclusion: Patchy and bilateral GGO along bronchi with predominant involvement of lower lungs are the most common chest CT findings of pandemic influenza A/H1N1 pneumonia.

• Clinical and Experimental Pediatrics
• /
• 제53권10호
• /
• pp.886-891
• /
• 2010

Purpose: This study aims to investigate the clinical characteristics of children diagnosed with the novel influenza A (H1N1) in the winter of 2009 at a single medical institution. Methods: Out of 545 confirmed cases of influenza A (H1N1) in children, using the real time RT-PCR method at Kosin University Gospel Hospital from September to December of 2009, 149 patients and their medical records were reviewed in terms of symptoms, laboratory findings, complications and transmission within a family. Results: Median age of subjects was 7 years (range: 2 months-18 years). New cases increased rapidly from September to reach a peak in November, then declined rapidly. Most frequently observed symptoms were fever (96.7%), cough (73.2%), rhinorrhea (36.9%) and sore throat (31.5%). Average body temperatures on the 1st, 2nd and 3rd hospital day were $38.75{\pm}0.65^{\circ}C$, $38.08{\pm}0.87^{\circ}C$ and $37.51{\pm}0.76^{\circ}C$, respectively. Complete blood counts and biochemical tests performed on the first admission day showed within the reference values in most cases. Of the 82 patients with simple chest radiography, 18 (22%) had pneumonic lesions; multi-focal bronchopneumonia in eleven, single or multi-segmental lobar pneumonia in five, and diffuse interstitial pneumonia in two patients. All of the 149 patients improved from their symptoms and discharged within 9 days of admission without any late complication. Conclusion: Children with 2009 pandemic influenza A (H1N1) at our single institution displayed nonspecific symptoms and laboratory findings, resembling those of common viral respiratory illnesses, and did not appear to develop more severe disease.

• 한국가금학회지
• /
• 제31권2호
• /
• pp.119-128
• /
• 2004

Highly Pathogenic Avian Influenza (HPAI) is a very acute systemic disease in poultry, particularly in chickens and turkeys caused by HPAI viruses. An outbreak of HPAI caused by subtype H5N1, was first reported in a broiler breeder farm on December 10, 2003 in Korea, although there had been twenty one outbreaks of the disease reported in the world before. Since mid-December 2003, eight Asian countries have confirmed outbreaks of HPAI due to the same subtype. The outbreak has also resulted in at least twenty three fatal human cases in Vietnam and Thailand as of May 17, 2004 according to the WHO. Regarding the first outbreak of recent Asian HPAI, it has been suspected that some Asian countries with the exception of Korea and Japan veiled the fact of HPAI outbreaks since the last half of 2003, even though it was first reported in Korea. There have been total nineteen outbreaks of HPAI among chicken and duck farms in 10 provinces in Korea since Dec. 2003 and approximately 5,280,000 birds were slaughtered from 392 farms for eradication of the disease and preemptive culling. The origin of the H5Nl HPAI virus introduced into the country are unknown and still under epidemiological investigation. Current status of outbreaks and characteristics of HPAI will be reviewed and discussed on the basis of genetic, virological, clinicopathological, and ecological aspect, as well as future measures for surveillance and prevention of the disease in Korea.

• Tuberculosis and Respiratory Diseases
• /
• 제72권6호
• /
• pp.493-500
• /
• 2012

Background: This study investigated the clinical characteristics and risk factors of the severity of pandemic influenza A (H1N1) 2009 infection in pediatric patients in Busan and Gyeongsangnam-do. Methods: Cases of influenza A (H1N1) 2009 in patients under the age of 18 years, confirmed by reverse transcription polymerase chain reaction, at Pusan National University Hospital and Pusan National University Yangsan Hospital from the last week of August 2009 through the last week of February 2010 were retrospectively analyzed. Results: Of the 3,777 confirmed cases of influenza A (H1N1) 2009, 2,200 (58.2%) were male and 1,577 (41.8%) were female. The average age of the patients was $8.4{\pm}4.8$ years. The total cases peaked during 44th to 46th week. Most of the patients were in the 5- to 9-year-old age group. Oseltamivir was administered to 2,959 (78.3%) of the patients. 221 patients (5.9%) were hospitalized, age an average of $6.7{\pm}4.5$ years. The average duration of hospitalization was $7.4{\pm}5.6$ days. One hundred cases (45.2%) had pneumonia. Risk factors for hospitalization included male gender, <2 years of age, and underlying disease. Children with asthma were at very high risk of hospitalization, over 20 times the non-asthmatic children (odds ratio [OR], 21.684; confidence interval [CI], 13.295~39.791). Likewise the children with neurologic deficits faced a 16 times higher risk (OR, 15.738; CI, 7.961~31.111). Ten of the patients (4.5%) were admitted to the intensive care unit, and eight (3.6%) required mechanical ventilation. Conclusion: Of the pediatric patients with pandemic influenza A (H1N1) 2009, most of the patients were in the 5- to 9-year-old age group. Risk factors for hospitalization included male gender, <2 years of age, and underlying disease. The most common complication was pneumonia. The very high risk of severe morbidity in children with asthma or neurologic disease shows the critical importance of targeted vaccine coverage, special awareness and swift care by both guardians and primary care providers.