• Journal of Microbiology and Biotechnology
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• v.34 no.2
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• pp.349-357
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• 2024

Foodborne pathogens, like Listeria monocytogenes, continue to inflict substantial financial losses on the food industry. Various methods for detecting Listeria in food have been developed and numerous studies have been conducted to compare the different methods. But, in recent years, new Listeria species have been identified, and currently the genus comprises 26 species. Therefore, it would be a more accurate approach to re-evaluate existing detection methods by considering new species. The present investigation involved the analysis of 42 ready-to-eat (RTE) foods, encompassing a variety of food categories, such as mezes, salads, dairy products, and meat products, with the aim of ascertaining the presence of Listeria. Among the traditional culture-dependent reference methods, the ISO 11290 method was preferred. The process of strain identification was conducted with the API Identification System. Furthermore, to ascertain the existence of L. monocytogenes and Listeria spp., the samples underwent additional analysis employing the VIDAS Immunoassay System, ELISA, and RT-PCR methodologies. Thus, four alternative approaches were employed in this study to compare not only the different methods used to determine Listeria while taking into account the newly identified Listeria species, but also to assess the compliance of retail RTE food items with microbiological criteria pertaining to the genus Listeria. Based on the conducted analyses, L. monocytogenes was conclusively determined to be present in one sample. The presence of Listeria spp. was detected in 30.9% of the samples, specifically in Turkish cig kofte, sliced salami, and salads.

• Biomedical Science Letters
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• v.17 no.4
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• pp.297-304
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• 2011

Influenza A virus of the Orthomyxoviridae family is a contagious respiratory pathogen that continues to evolve and burden in the human public health. It is able to spread efficiently from human to human and have the potential to cause pandemics with significant morbidity and mortality. It has been estimated that every year about 500 million people are infected with this virus, causing about approximately 0.25 to 0.5 million people deaths worldwide. Influenza A viruses are classified into different subtypes by antigenicity based on their hemagglutinin (HA) and neuraminidase (NA) proteins. The sudden emergence of influenza A virus subtypes and access for epidemiological analysis of this subtypes demanded a rapid development of specific diagnostic tools. Also, rapid identification of the subtypes can help to determine the antiviral treatment, because the different subtypes have a different antiviral drug resistance patterns. In this study, our aim is to detect influenza A virus subtypes by using real-time nucleic acid sequence based amplification (NASBA) which has high sensitivity and specificity through molecular beacon. Real-time NASBA is a method that able to shorten the time compare to other molecular diagnostic tools and is performed by isothermal condition. We selected major pandemic influenza A virus subtypes, H3N2 and H5N1. Three influenza A virus gene fragments such as HA, NA and matrix protein (M) gene were targeted. M gene is distinguished influenza A virus from other influenza virus. We designed specific primers and molecular beacons for HA, NA and M gene, respectively. In brief, the results showed that the specificity of the real-time NASBA was higher than reverse transcription polymerase chain reaction (RT-PCR). In addition, time to positivity (TTP) of this method was shorter than real-time PCR. This study suggests that the rapid detection of neo-appearance pandemic influenza A virus using real-time NASBA has the potential to determine the subtypes.

• Journal of Life Science
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• v.21 no.6
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• pp.819-828
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• 2011

To inspect norovirus contamination of groundwater in south eastern areas of Korea, a systematic survey of groundwater in Busan, Ulsan, and Gyeongsangnam-do was performed for two years from 2009 to 2010. For this purpose, we first optimized the nested reverse transcription-PCR condition by designing two sets of primers for the detection of norovirus genogroups, GI and GII. Of 145 samples, 21 (25.9%) and 15 (23.4%) were norovirus positive in the dry season (April to June) and wet season (July to August), respectively. The detection frequencies of norovirus in Busan, Ulsan, and Gyeongsangnam-do were 15%, 7%, and 32%, respectively, reflecting a geographical difference in their distribution. The GI and GII isolates were 5 and 31, respectively, indicating the prevalence of GII in the tested areas. According to phylogenetic analysis of their nucleotide sequences, all of the GI isolates were identified to genotype GI.5 whilst the GII isolates were divided into two genotypes, GII.3 and GII.4. Neither physical-chemical parameters such as pH, temperature, oxidation-reduction potential, and dissolved oxygen, nor microbial indicators of water quality such as total bacteria, total coliforms, and Escherichia coli were statistically correlated with contamination of norovirus in the groundwater. Interestingly, however, the presence of norovirus was closely correlated with low turbidity (<0.50 NTU). The present study suggests that periodical monitoring of norovirus in groundwater is necessary to prevent epidemic waterborne diseases and to secure better sanitary conditions for public health.

• The Plant Pathology Journal
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• v.26 no.2
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• pp.130-138
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• 2010

Orchid fleck virus (OFV) is an unassigned plant virus in the family Rhabdoviridae. OFV was isolated from Cymbidium sp. showing oval necrotic lesions on their leaves in Korea, and designated as OFV-NHHS1. The complete nucleotide sequence of the RNA1 (6,413 nt) (GenBank accession no. AB516442) and RNA2 (6,001 nt) (GenBank accession no. AB516441) was determined in this study. RNA1 and RNA2 contained five and one ORF respectively. RNA1 encodes nucleocapsid (N) of 49 kDa, ORF2 of 26 kDa, ORF3 of 38 kDa, ORF4 of 20 kDa and glycoprotein (G) of 61 kDa proteins, whereas RNA2 encodes a single polymerase of 212 kDa. OFV-NHHS1 shared extremely high similarity of 98.6-100% and 98.9-99.6% in nucleotidle and amino acid sequences with a Japanese isolate, OFV-so, respectively. However, the N, G and L of OFV-NHHS1 revealed 6.9-19.3%, 7.3-12.0%, and 13.4-26.6% identities to those of 29 Rhabdoviruses, respectively. To survey the infection rate of OFV in commercial orchids in Korea, 51 Cymbidium sp., 10 Phalaenopsis sp., 22 Oncidium sp. and 21 Dendrobium sp. plants that showed typical viral symptoms were collected. RT-PCR with specific primers for detection of Cymbidium mosaic virus (CymMV), ORSV and OFV showed high infection rate by ORSV alone and double infection by ORSV and CymMV. One of the orchids tested was infected with OFV. This is the first report of the complete nucleotide sequences of OFV isolated in Korea.

• Korean Journal of Veterinary Research
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• v.37 no.4
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• pp.793-807
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• 1997

We tried to develop detection system of porcine reproductive and respiratory syndrome virus(PRRSV) by in situ hybridization(ISH) in the piglets experimentally infected with KPRRS-2, the Korean isolate(12 piglets) or Mn-1b, the American isolate(4 piglets), and in the natural infection suspected 6 piglets. Twelve 30-days-old piglets(two pigs per each inoculated group) were inoculated by nasal instillation of KPRRS-2 virus(total dose $10^{4.5}TCID_{50}$), Six piglets(one pig per each group) were induced contact infection with inoculated piglets, during the experiment, and two piglets were used as control. Inoculated or contacted piglets were euthanized at 1, 3, 5, 7, 14 and 21 days postinoculation(DPI). The respiratory signs such as coughing and nasal discharge were observed on day 3 DPI, and ear cyanosis were on day 5 DPI, including contacted piglets. Through the necropsy, purple discolorization of dorsal part of lung, and hypertrophy of local lymph nodes were observed. The histopathological lesions of lung were interstitial pneumonia characterized by type 2 pneumocyte hyperplasia. We prepared the probe for ISH by RNA isolation from KPRRS-2, RT-PCR, and biotin labeling. We performed the ISH within only 1~2 hours using $Microprobe^{TM}$ capillary action system. As the results, the strong red specific positive signals, means PRRSV distribution, was mainly observed in the cytoplasm of alveolar macrophages. And also signals were detected in some type 2 pneumocytes and bronchiolar epithelium of lung, myocardium, liver, kidney, tonsil, spleen, gastrointestinal mucosa, testis and lymph nodes.

• Journal of Environmental Health Sciences
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• v.38 no.1
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• pp.57-65
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• 2012

Objectives: For our survey of the incidence of norovirus infections and the genogroup distribution of norovirus in Seoul, Republic of Korea, we evaluated through regular surveillance the prevalence of norovirus infections in patients with acute gastroenteritis occurring in Seoul from January 2007 to July 2011. Methods: For norovirus detection, we conducted epidemiological analyses on the basis of the junction of ORF1 and ORF2 (approximately 314 bp). 11,202 fecal specimens were collected from patients in Seoul with acute gastroenteritis between January 2007 and July 2011 and then tested for the presence of NoV via reverse transcription (RT) - polymerase chain reaction (PCR). Results: 16.6% (1,861/11,202) of the fecal specimens were determined to be positive for noroviruses. The incidences of norovirus infection in Seoul in the case of acute gastroenteritis with regular surveillance were 28.0% in 2007, 14.6% in 2008, 9.1% in 2009, 14.1% in 2010, and 12.9% in 2011, which shows that noroviruses constituted a major causative agent of acute gastroenteritis. Also, the incidence of noroviral infection in patients with acute gastroenteritis increased after the large-scale new influenza in 2009. Conclusions: The genetic characteristics of norovirus and the epidemiologic patterns of a viral pathogen in acute gastroenteritis patients may provide potentially effective data for epidemiological studies in Seoul, Korea.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.2
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• pp.109-115
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• 2016

The efficacy of depuration following growing area translocation for the defecation of norovirus was evaluated under experimental conditions using oysters Crassostrea gigas previously subjected to bioaccumulation of this virus at a waste treatment plant discharge site. Three trials were assayed in an open experimental system with a commercial oyster farm located in a shellfish growing area approved by the Korean Shellfish Sanitation Program. Real-time reverse-transcription polymerase chain reaction (RT-PCR) was used to quantify viruses in the digestive glands of oysters. The final viral loads in oysters after 12 days remained under the detection limit (10 copies/g digestive gland) of the real-time RT-PCR. This reduction trend showed two-phase removal kinetics, with an initial slow reduction or slight increase in viruses during the first 2 days of depuration and subsequent stabilization with 0.12 to 2.64 log unit norovirus copies/g digestive gland per 2 days of depuration for the remaining time.

• Research in Plant Disease
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• v.15 no.1
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• pp.8-12
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• 2009

Virus disease surveys of strawberries cultivated and preserved as germplasm resources in Korea was conducted during 2007-2008. Virus detection was conducted by RT-PCR using total RNAs extracted from strawberry samples. We detected the infection with Strawberry mild yellow edge virus (SMYEV), Strawberry mottle virus (SMoV), Strawberry vein banding virus (SVBV) and Strawberry pallidosis associated virus (SPaV) while no infection with Strawberry crinkle virus (SCV), Strawberry necrotic shock virus (SNSV), Strawberry latent ring spot virus (SLRSV) and Arabis mosaic virus (ArMV) was observed. The infection rate of virus disease on 4 cultivars including Seolhyang, Maehyang, Gumhyang, and Dahong, bred in Korea, was 0.1, 1.9, 0, and 0%, respectively. Surprisingly, however, cultivar Red Peal introduced from Japan in 1997 revealed 48.3% virus infection rate. SMYEV, SMoV and SPaV were also identified in strawberries growing in the farm fields of Korea. In the field, however, SMYEV was the most predominant virus (97.4%) among those 3 identified viruses. SVBV was detected only in strawberry kept as a germplasm.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.4
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• pp.1705-1712
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• 2016

Background: The aim of this study was to determine and evaluate the association of different viral infections, with hepatitis B and C viruses, Epstein-Barr virus, cytomegalovirus and human herpes virus-8 (HBV, HCV, EBV, CMV, HHV-8) with the risk of lymphomas (Hodgkin and non-Hodgkin) among Egyptian patients, and correlate with the histopathological staging and typing as well as the prevalence of combined infections. Materials and Methods: A total of 100 newly diagnosed lymphoma patients with 100 healthy age and sex matched normal controls were assayed for viral infection using enzyme linked immunosorbant assay (ELISA) followed by real time polymerase chain reaction (RT-PCR). Results: Our results showed a high statistical significant difference between cases and controls as regards clinical and laboratory findings (P<0.001 and=0.003). A high statistical difference was seen for the association of most viruses and lymphoma cases (p<0.001) except for positive HBs Ag, positive CMV IgG and HHV-8 (p=0.37, 0.70 and 1.0 respectively). No statistical significant difference was found between Hodgkin (HL) and non-Hodgkin (NHL) as regards viral prevalence except HCV antigen, 57.1% for HL and 26.5% for NHL (p = 0.03). Only, HBV DNA showed a high significant value among infiltrated bone marrow cases (p=0.003) and finally, a high significant association of 2 combined viral infections with infiltrated bone marrow lymphoma cases (p=0.04). Conclusions: Our results showed that infection with HBV, HCV, CMV and EBV were associated with increased risk of lymphoma among the Egyptian population. Detection of new associations between infectious agents and risk of cancer development will facilitate progress in elaboration of prophylactic measures, early diagnostic methods and, hopefully, novel therapy of malignant tumours.

• Korean Journal of Veterinary Service
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• v.38 no.1
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• pp.1-7
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• 2015

This study was investigated to diagnose pathogenic organisms of atrophic piglets in northern area of the Gyeongnam province, Korea. Samples such as feces, blood and necropsy specimens of 42 atrophic piglets (${\leq}10$ weeks old) were taken from May to December 2013 for this survey. Samples were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) assay and bacteria isolation for detection of pathogenic agents. 93 pathogens were isolated from 42 samples can be classified into ll groups. We identified bacterial agents in 56 cases (60.2%) and viral agents in 31 cases (33.3%). However, 6 cases (6.5%) were undetected. Among these pathogens, the most prevalent disease were porcine reproductive and respiratory syndrome (PRRS) in 22 cases (23.7%). The major diseases were Colibacillosis in 15 cases (16.1%), Glasser's disease in 12 cases (12.9%), and porcine epidemic disease (PED) in 9 cases (9.7%). Mixed infections were accounted for 77.8% of atrophic piglets. In particular, the rate of mixed infections with PRRS virus showed the highest frequency (71.4%). In addition, there is a seasonal variation. Viral pathogens were dominantly detected in winter, but in the rest of the season bacterial agents were mainly detected. Gastrointestinal diseases occurred mainly in the pre-weaning piglets, the respiratory diseases and wasting diseases occurred mainly in the post-weaning piglets.