• Korean Journal of Veterinary Service
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• v.40 no.1
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• pp.15-20
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• 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.

• Journal of The Korean Society of Agricultural Engineers
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• v.53 no.1
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• pp.29-36
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• 2011

HPAI (High pathogenic avian influenza) which is a disease legally designated as an epidemic generally shows rapid spread of disease resulting in high mortality rate as well as severe economic damages. Because Korea is contiguous with China and southeast Asia where HPAI have occurred frequently, there is a high risk for HPAI outbreak. A prompt treatment against epidemics is most important for prevention of disease spread. The spread of HPAI should be considered by both direct and indirect contact as well as various spread factors including airborne spread. There are high risk of rapid propagation of HPAI flowing through the air because of collective farms mostly in Korea. Field experiments for the mechanism of disease spread have limitations such as unstable weather condition and difficulties in maintaining experimental conditions. In this study, therefore, computational fluid dynamics which has been actively used for mass transfer modeling were adapted. Korea has complex terrains and many livestock farms are located in the mountain regions. GIS numerical map was used to estimate spreads of virus attached aerosol by means of designing three dimensional complicated geometry including farm location, road network, related facilities. This can be used as back data in order to take preventive measures against HPAI occurrence and spread.

• Korean Journal of Veterinary Service
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• v.42 no.3
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• pp.135-144
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• 2019

Low pathogenic avian influenza (LPAI; H9N2) and Newcastle disease (ND) are economically important poultry diseases in Korea. In this study, we investigated pathological features and virus distribution in the lymphoid tissues of chicks experimentally infected with H9N2 and/or ND virus. Six-weeks-old SPF chickens were divided into 4 groups, Control (C), H9N2 (E1), NDV (E2), and H9N2+NDV (E3). E1 group was challenged with 0.1 ml A/Kr/Ck/01310/01 (H9N2) $10^{5.6}$ $EID_{50}$ intranasally, E2 group was challenged with 0.5 ml KJW (NDV) $10^{5.0}{\sim}10^{6.0}$ $ELD_{50}$ intramuscularly, and E3 group was challenged with H9N2, followed 7 days later by NDV. In histopathological examination, E1 group showed depletion and necrosis in bursa of Fabricius, thymus, cecal tonsil, and spleen, whereas E2 and E3 groups were noted severe lymphocyte depletion and necrosis with destruction of lymphoid organs structures. In TUNEL assay, apoptotic bodies were detected in lymphoid organs of all experimental groups, which was most severe in E3 group. H9N2 and ND viruses were predominantly detected in cecal tonsil of E1, E2, and E3 groups by PCR and immunohistochemistry (ICH). In conclusion, co-infection of H9N2 with NDV caused severe pathologic lesions and apoptosis in lymphoid tissues compared to single infections.

• Korean Journal of Pharmacognosy
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• v.30 no.1
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• pp.25-33
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• 1999

In order to find less toxic antiviral agents from basidiomycetes, EA, the water soluble substance, was prepared from the carpophores of Elfvingia applanata (Pers.) Karst. EA was examined for antiviral activity against five strains of pathogenic viruses such as encephalomyocarditis virus (EMCV), vesicular stomatitis virus (VSV) Indiana and New Jersey strains, influenza A virus (Flu A), and varicella zoster virus (VZV) in vitro. Antiviral activity was evaluated by plaque reduction assay. Among five strains of viruses tested, EA exhibited the most potent antiviral activity against VSV Indiana strain with 50% effective concentration $(EC_{50})$ of 0.104 mg/ml in Vero cells, and its selectivity index (SI) was 36.5. EA was also examined for the virucidal activity, antiviral activity in preincubation on VSV Indiana strain in order to examine possible mode of antiviral activity. Preincubation of Vero cells with EA did not confer protection against VSV, however, prolonged exposure of cells to EA inhibited the replication of virus dose-dependently. In virucidal activity, the titer of infectious virus did not decrease significantly.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.178-185
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• 2008

Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.

• Tuberculosis and Respiratory Diseases
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• v.68 no.4
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• pp.205-211
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• 2010

Background: Procalcitonin is a well known marker in infection that plays a role in distinguishing between bacterial and viral infections in screening. The aim of the present study was to evaluate the role of procalcitonin in differentiating between 2009 H1N1 influenza pneumonia and community acquired pneumonia of bacterial origin, or mixed bacterial origin and 2009 H1N1 influenza infection. Methods: A retrospective observational study was performed over the 6-month winter period during the 2009 H1N1 influenza pandemic. Ninety-six patient-subjects were enrolled, all of whom had been diagnosed with community acquired pneumonia in emergency department during the study period. On admission, laboratory studies were performed, which included 2009 H1N1 influenza real-time polymerase chain reaction of nasal secretions and procalcitonin on serum; the laboratory values were compared between the study groups. Receiver operating characteristic curve analyses were performed on the resulting data. Results: Compared to those with bacterial or mixed infections (n=62) and bacterial pneumonia with confirmed organisms (n=30), patients with 2009 H1N1 pneumonia (n=34) were significantly more likely to have low procalcitonin levels (p=0.008, 0.001). Using cutoff of value >0.3 ng/mL, the sensitivity and specificity of procalcitonin for detection of patients with confirmed bacterial pneumonia were 76.2% and 60.6%, respectively. A significant difference in procalcitonin was found between 2009 H1N1 pneumonia and pneumonia caused by mixed influenza viral and bacterial infections (0.15 [0.05~0.84] vs. 10.3 [0.05~22.87] ng/mL, p=0.045). Conclusion: Serum procalcitonin measurement may assist in the discrimination between pneumonia of bacterial and of 2009 H1N1 influenza origin. High values of procalcitonin suggest that bacterial infection or mixed infection of bacteria and 2009 H1N1 influenza is more likely.

• Tuberculosis and Respiratory Diseases
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• v.59 no.3
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• pp.279-285
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• 2005

Background : Ambient particles during Asian dust events are usually less than $10{\mu}m$ in size, and known to be associated with the adverse effects on the general population. There is little evidence linking Asian dust to adverse effects on the airways. In 2002, the authors found that particulate matter during Asian dust events had an effect on the symptoms and pulmonary function of patients with bronchial asthma. An aggravating factor might be that of a viral infection, but this remains unclear. Conversely, it has been speculated that African dust may carry the virus responsible for foot and mouth disease. Asian dust events are also likely to be responsible for transporting viruses, some of which are pathogenic, and common in many environments. Therefore, in this study, air samples were screened for the presence of viruses. Methods : Air samples were collected 20 times each during Asian dust events and under non-dust conditions, for at least 6 hours per sample, using a high volume air sampler (Sibata Model HV500F), with an airflow rate of 500L/min, between April and August 2003, and between April and August 2004. The samples were then screened for the presence of targeted viruses (Influenza A, B, Hog cholera virus, and Aphthovirus) using a polymerase chain reaction method. Results : One Asian dust event occurred between April and August 2003, and 3 between April and August 2004, with a 24 hour average PM10 level of $148.0{\mu}g/m^3$. The 24 hour average PM10 level was $57{\mu}g/m^3$. There was a significant difference in the PM10 concentration between dusty and clear days. No viruses (Influenza virus, Aphthovirus, and Hog cholera virus) were identified in the air samples obtained during the dusty days. Conclusions : Although no virus was detected in this study, further studies will be needed to identify suspected viruses carried during Asian dust events, employing more appropriate virus detection conditions.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.4
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• pp.465-470
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• 2009

The chicken Mx gene has been regarded as a candidate gene for resistance to avian influenza virus (AIV). In this study, three groups of chickens with homozygotes (AA, GG) and heterozygotes (AG) of the resistant (A) and susceptible alleles (G) to AIV of the Mx gene were constructed from a line of dwarf egg-type chickens. These chickens were not examined for their resistant activities to AIV because the differential resistance had only been detected in vitro. The birds of the three groups were vaccinated with inactivated H5N2 AIV vaccine and the level of hemagglutination inhibition (HI) antibody to AIV was detected. The association between disease resistant activity to AIV and antibody response to AIV vaccination in the three groups was analyzed. The chickens with homozygous resistant allele A showed the lowest antibody levels, whereas the heterozygous chickens (AG) presented the highest antibody level after the boosting vaccination, which indicates that the efficiency of artificial selection on the resistant allele of Mx gene will be compromised since the homozygotes of the allele presented the weakest antibody response to the corresponding vaccine.

• Biomedical Science Letters
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• v.6 no.1
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• pp.19-28
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• 2000

This study was performed to evaluate the activities of glutathione S-transferase (GST) and lactate dehydrogenase (LDH) in Maddin-Darby canine kidney (MDCK) cells infected with virus and/or treated with amantadine. On cell morphological findings, monolayer fractions in MDCK cells infected with virus were exfolated more than 80% in 1 TCID$_{50}$ group and that in 10 TCID$_{50}$ were completely exfolated after 3 days during infectious process. In proportion to the dose of amantadine, activities of GST and LDH of MDCK cells were significantly decreased and those of LDH in medium fraction were more significantly increased compared with control. According to in both dose and time of virus innoculation, activities of GST and LDH in MDCK cells were significantly decreased in 1 and 10 TCID$_{50}$ infected cells after 3 days. LDH activities in infectious medium were remarkably rised at 10 fold. In case of the cell line inoculated with type A 100 TCID$_{50}$ and additionally treated with amantadine, the decreasing rate to the control in activities of GST and LDH was lower than that in those in case of that infected with virus only. These results suggested that virus infection and amantadine treatment may effect the activity of the detoxicating enzyme in the target cells.

• Genomics & Informatics
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• v.18 no.4
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• pp.40.1-40.8
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• 2020

Avian influenza (AIV) outbreaks can induce fatal human pulmonary infections in addition to economic losses to the poultry industry. In this study, we aimed to develop a rapid and sensitive point-of-care AIV test using loop-mediated isothermal amplification (LAMP) technology. We designed three sets of reverse transcription LAMP (RT-LAMP) primers targeting the matrix (M) and hemagglutinin (HA) genes of the H5 and H9 subtypes. RT-LAMP targeting the universal M gene was designed to screen for the presence of AIV and RT-LAMP assays targeting H5-HA and H9-HA were designed to discriminate between the H5 and H9 subtypes. All three RT-LAMP assays showed specific amplification results without nonspecific reactions. In terms of sensitivity, the detection limits of our RT-LAMP assays were 100 to 1,000 RNA copies per reaction, which were 10 times more sensitive than the detection limits of the reference reverse-transcription polymerase chain reaction (RT-PCR) (1,000 to 10,000 RNA copies per reaction). The reaction time of our RT-LAMP assays was less than 30 min, which was approximately four times quicker than that of conventional RT-PCR. Altogether, these assays successfully detected the existence of AIV and discriminated between the H5 or H9 subtypes with higher sensitivity and less time than the conventional RT-PCR assay.