• Title/Summary/Keyword: multiplex polymerase chain reaction (PCR)

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The Efficacy of Rapid Antigen Tests for Detection of Seasonal Influenza Virus (계절 인플루엔자 진단에서 신속항원검사의 유용성)

  • Lee, Jun Yeol;Lee, Sol;Kim, Han Sung;Kim, Kwang Nam
    • Pediatric Infection and Vaccine
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    • v.24 no.1
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    • pp.31-36
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    • 2017
  • Purpose: This study aimed to examine the accuracy of rapid influenza diagnostic tests (RIDT) in children with an influenza-like illness and to evaluate factors associated with greater accuracy. Methods: Pediatric patients, who visited Hallym University Sacred Heart Hospital with an influenza-like illness between June 2011 and May 2016, were enrolled in this study. We tested 798 samples using a real-time polymerase chain reaction (PCR) for respiratory viruses and compared the results with rapid influenza tests. Results: In comparison with the results of the multiplex PCR, the positive agreement rates of RIDT for influenza A and B virus were 75.7% and 60.0%, respectively. The performance of RIDT varied according to days after fever onset. The positive agreement rates of RIDT for influenza A and B tests, performed within 4 days of fever onset, were 77.6% and 73.2%, but the rates for tests performed more than 5 days after fever onset were 66.7% and 21.4%, respectively. Conclusions: The RIDT is a quick and simple aid to diagnosis, but is less sensitive than the labeled sensitivity. Moreover, test performance varied according to days after fever onset. Test specimens for RIDT should be collected as soon as possible after the onset of symptoms (less than 4 days).

Clinical Characteristics and Ultrasonographic Findings of Acute Bacterial Enterocolitis in Children

  • Chun, Peter;Lim, Taek Jin;Hwang, Eun Ha;Mun, Sang Wook;Lee, Yeoun Joo;Park, Jae Hong
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.20 no.2
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    • pp.107-113
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    • 2017
  • Purpose: This study clarified the bacterial pathogens currently causing acute infectious enterocolitis (AIE) in children and evaluated the clinical characteristics and ultrasonographic findings according to the different pathogens. Methods: Medical records regarding age, sex, clinical symptoms, laboratory data, identified enteropathogens, ultrasonographic findings, treatment, and outcome of 34 patients who were diagnosed with AIE via stool examination using multiplex polymerase chain reaction (PCR) or culture, were retrospectively reviewed. Results: Twenty-four patients (70.6%) were male. The mean age of the patients was $8.5{\pm}6.2$ (range, 1.1-17.1) years. Six bacterial pathogens were isolated: Salmonella species (spp.) (32.4%), Campylobacter spp. (20.6%), verotoxin-producing Escherichia coli (14.7%), Staphylococcus aureus (11.8%), Clostridium difficile (8.8%), and Shigella spp. (2.9%). Abdominal pain occurred in all patients regardless of pathogen. The patients infected with Salmonella were older than those infected with verotoxin-producing E. coli (p<0.05). C-reactive protein levels were higher in patients with Salmonella and Campylobacter infections than in those with verotoxin-producing E. coli infection (p< 0.05), the other clinical and laboratory data were indistinguishable between pathogens. Ultrasonography demonstrated diverse involvement of bowel segments according to pathogen. Wall thickening of both the ileum and the entire colon was the most common lesion site regardless of pathogen. Conclusion: Various bacterial agents cause AIE and the symptoms are diverse symptoms, however, all most children recovered spontaneously. Use of multiplex PCR on stool samples warrants improvement of its sensitivity for diagnosis of enteropathogenic bacteria. Ultrasonographic examination is useful for diagnosis of AIE; it can also detect the disease extent and severity.

Respiratory Syncytial Virus Outbreak in the Basic Military Training Camp of the Republic of Korea Air Force

  • Park, Won-Ju;Yoo, Seok-Ju;Lee, Suk-Ho;Chung, Jae-Woo;Jang, Keun-Ho;Moon, Jai-Dong
    • Journal of Preventive Medicine and Public Health
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    • v.48 no.1
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    • pp.10-17
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    • 2015
  • Objectives: An outbreak of acute febrile illness occurred in the Republic of Korea Air Force boot camp from May to July 2011. An epidemiological investigation of the causative agent, which was of a highly infective nature, was conducted. Methods: Throat swabs were carried out and a multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) assay was performed to identify possible causative factors. Results: The mean age of patients who had febrile illness during the study period was 20.24 years. The multiplex RT-PCR assay identified respiratory syncytial virus (RSV) as the causative agent. The main symptoms were sore throat (76.0%), sputum (72.8%), cough (72.1%), tonsillar hypertrophy (67.9%), and rhinorrhea (55.9%). The mean temperature was $38.75^{\circ}C$ and the attack rate among the recruits was 15.7% (588 out of 3750 recruits), while the mean duration of fever was 2.3 days. The prognosis was generally favorable with supportive care but recurrent fever occurred in 10.1% of the patients within a month. Conclusions: This is the first epidemiological study of an RSV outbreak that developed in a healthy young adult group. In the event of an outbreak of an acute febrile illness of a highly infective nature in facilities used by a young adult group, RSV should be considered among the possible causative agents.

Discrimination of Korean ginseng (Panax ginseng Meyer) cultivar Chunpoong and American ginseng (Panax quinquefolius) using the auxin repressed protein gene

  • Kim, Jong-Hak;Kim, Min-Kyeoung;Wang, Hongtao;Lee, Hee-Nyeong;Jin, Chi-Gyu;Kwon, Woo-Saeng;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.395-399
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    • 2016
  • Background: Korean ginseng (Panax ginseng) is one of the most important medicinal plants in the Orient. Among nine cultivars of P. ginseng, Chunpoong commands a much greater market value and has been planted widely in Korea. Chunpoong has superior quality "Chunsam" ($1^{st}$ grade ginseng) when made into red ginseng. Methods: A rapid and reliable method for discriminating the Chunpoong cultivar was developed by exploiting a single nucleotide polymorphism (SNP) in the auxin repressed protein gene of nine Korean ginseng cultivars using specific primers. Results: An SNP was detected between Chunpoong and other cultivars, and modified allele-specific primers were designed from this SNP site to specifically identify the Chunpoong cultivar and P. quinquefolius via multiplex polymerase chain reaction (PCR). Conclusion: These results suggest that great impact to prevent authentication of precise Chunpoong and other cultivars using the auxin repressed protein gene. We therefore present an effective method for the authentication of the Chunpoong cultivar of P. ginseng and P. quinquefolius.

Selective Detection of Campylobacter jejuni, C. coli, Arcobacter butzleri and Helicobacter pylori by Polymerase Chain Reaction (Campylobacter jejuni, C. coli, Arcobacter butzleri와 Helicobacter pylori의 PCR에 의한 분리검출)

  • Lee, Young-Duck;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.34 no.6
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    • pp.1134-1139
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    • 2002
  • Campylobacter, Arcobacter, and Helicobacter, classified into the same rRNA superfamily VI by taxonomy, cause food-borne diseases, stomach ulcer, and gastric cancer. To detect each strain selectively from contaminated foods, PCR, multiplex-PCR, and restricion fragment length polymorphism (RFLP) were applied on Campylobacter, Arcobacter, and Helicobacter. The same PCR products could be detected using CHA primer targeted for 16S rRNA of Campylobacter, Arcobacter, and Helicobacter. To detect C. jejuni and C. coli from A. butzleri and H. pylori, pg50/pg3 primer targeted for fla A gene was used, and for A. butzleri, Arco2/Butz primer targeted for 23S rRNA was utilized. For H. pylori detection, icd1/icd2 primer targeted for isocitrate dehydrogenase gene was employed, and JEJ1/JEJ2 primer targeted for ceuE gene was effective for C. jejuni detection from the three strains. C. jejuni, C. coli could be separated from A. butzleri and H. pylori through PCR-RFLP using restriction enzyme Dde I. Such primers would be effective for detecting each strain selectively through PCR when C. jejuni, C. coli, A butzleri and H. pylori are contaminated together.

Recent (2010-2019) foodborne outbreaks caused by viruses in the Republic of Korea along with their detection and inactivation methods (바이러스에 의한 최근(2010-2019) 국내 식중독 사고와 검출법 및 제어법에 대한 동향 조사)

  • Kwon, Seung-Wook;Kim, Sang-Soon
    • Korean Journal of Food Science and Technology
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    • v.53 no.1
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    • pp.1-11
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    • 2021
  • In this review, recent foodborne outbreaks caused by viruses in the Republic of Korea (2010-2019) were analyzed. The human norovirus was found to be the major foodborne virus causing an average of 94.9% of the viral outbreaks. Reverse-transcription polymerase chain reaction (PCR) with electrophoresis has been widely used to detect viruses, but several rapid detection methods, including real-time PCR, multiplex PCR, and quantum dot assay, have also been suggested. For norovirus inactivation studies, surrogates such as murine norovirus and feline calicivirus have been widely used to identify the reduction rate owing to the limitations in laboratory cultivation. Conversely, direct cell infection studies have been conducted for other foodborne viruses such as adenovirus, astrovirus, rotavirus, and hepatitis A or E virus. Moreover, virucidal mechanisms using various physical and chemical treatments have been revealed. These recent studies suggest that rapid in situ detection and effective control are valuable for ensuring food safety against viral infections.

Chlamydia trachomatis respiratory infection in Korean young infants (국내 영유아에서의 Chlamydia trachomatis 호흡기 감염)

  • Hong, Ki Bae;Shin, Youn Shim;Roh, Eui-Jung;Chung, Eun Hee
    • Clinical and Experimental Pediatrics
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    • v.51 no.7
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    • pp.729-735
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    • 2008
  • Purpose : Chlamydia trachomatis is one of the most common sexually transmitted diseases and is also a cause of pneumonia in infants. Respiratory infections by respiratory viruses are also common for infants. The objectives of this study were to identify the clinical manifestations and to determine the prevalence of C. trachomatis respiratory infections and coinfections by respiratory viruses in infants younger than 6 months of age. Methods : For this study, we enrolled 6 months or younger infants who were admitted to the Dankook University Hospital between January 2002 and July 2007, with respiratory symptoms. Nasopharyngeal aspirates or throat swabs were collected within s d of hospitalization and C. trachomatis was detected using polymerase chain reaction (PCR). Patients who tested positive underwent multiplex PCR for respiratory viruses. Results : A total of 690 patients underwent chlamydial PCR testing and 36 (5.2%) had positive results. Of the 36, 28 (78%) were male; 30 were vaginally delivered. From the 36 patients positive for C. trachomatis, 26 underwent multiplex respiratory viral PCR; 12 were coinfected with viruses. Respiratory syncytial virus (RSV) was the most frequent pathogen that was detected in 6 patients. Increased C-reactive protein and fever were significant in patients coinfected with respiratory viruses. Conclusion : C. trachomatis can infected in infants delivered by cesarean section as well as in 6 months old or younger infants. Infant with C. trachomatis respiratory infections can also be coinfected with respiratory infection also coinfected with respiratory viruses. Further studies are needed to better understand the prevalence rates of the this infection and its coinfection rate with respiratory viruses.

Occurrence of Apple stem grooving virus in commercial apple seedlings and analysis of its coat protein sequence

  • Han, Jae-Yeong;Park, Chan-Hwan;Seo, Eun-Yeong;Kim, Jung-Kyu;Hammond, John;Lim, Hyoun-Sub
    • Korean Journal of Agricultural Science
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    • v.43 no.1
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    • pp.21-27
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    • 2016
  • Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV), and Apple stem pitting virus (ASPV) have been known to induce top working disease causing economical damage in apple. Occurrences of these three viruses in pome fruit trees, including apple, have been reported around the world. The transmission of the three viruses was reported by grafting, and there was no report of transmission through mechanical contact, insect vector, or seed except some herbaceous hosts of ASGV. As RNA extraction methods for fruit trees, Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and multiplex RT-PCR techniques have been improved for reliability and stability, and low titer viruses that could not be detected in the past have become detectable. We studied the seed transmission ability of three apple viruses through apple seedling diagnosis using RT-PCR. Nineteen seeds obtained from commercially grown apple were germinated and two of the resulting plants were ASGV positive. Seven clones of the amplified ASGV coat protein (CP) genes of these isolates were sequenced. Overall sequence identities were 99.84% (nucleotide) and 99.76% (amino acid). Presence of a previously unreported single nucleotide and amino acid variation conserved in all of these clones suggests a possible association with seed transmission of these 'S' isolates. A phylogenetic tree constructed using ASGV CP nucleotide sequences showed that isolate S sequences were grouped with Korean, Chinese, Indian isolates from apple and Indian isolates from kiwi.

Detection of beta-lactam antibiotic resistant genes in Escherichia coli from porcine fecal samples using DNA chip

  • Park, Nam-Yong;Na, Sung-Ho;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.30 no.4
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    • pp.505-510
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    • 2007
  • This study was conducted to detect ${\beta}$-lactam antibiotic-resistant genes in the 400 E coli isolates from porcine fecal samples in Korea by a DNA chip. The DNA chip contains the specific probe DNAs of the ${\beta}$-lactam antibiotic-resistant genes that had been labeled with a mixture of primer set designed to amplify specific genes (PSE, OXA, FOX, MEN, CMY, TEM, SHV, OXY and AmpC) using a multiplex polymerase chain reaction (PCR). Of 400 isolates 339 contained at least one ${\beta}$-lactamases gene. Resistance to ${\beta}$-lactamases was mediated mainly by AmpC (n = 339, 100%), and followed by TEM (n = 200, 59.0%), CMY (n = 101, 29.8%), PSE (n = 30, 8.9%) and both OXA and SHV genes (n = 20, 5.9%), while the FOX, MEN and OXY genes were not detected. The other sixty-one did not contain any ${\beta}$-lactamase genes even though they were resistant to antimicrobial drugs. In conclusion, the DNA chip system can be used as a rapid and reliable method for detecting of ${\beta}$-lactamases genes, which will help veterinarians select the antibiotics for monitoring and treating of animal diseases.

Characteristics and Pathogenicity for Japnaes Eel Anguilla japonica of Vibrio vulnificus Isolated from Oyster, Sediment and Seawater in the Korea Coast

  • Kim, Myoung Sug;Jung, Sung Hee;Hong, Suhee;Jeong, Hyun Do
    • Fisheries and Aquatic Sciences
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    • v.18 no.4
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    • pp.387-393
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    • 2015
  • Biotyping of Vibrio vulnificus strains isolated from marine environments along the south coast of Korea showed that the majority of the isolates (94.7%) belonged to biotype 1 and the remaining isolates (5.3%) belonged to biotype 2. Analysis of 16S rRNA V. vulnificus strains isolated from marine environments using a multiplex polymerase chain reaction (PCR) revealed that 78.7% were type A and 21.3% were type B. Random amplified polymorphic DNA (RAPD) was used to analyze the genomic differences in V. vulnificus among the biotype 2 strains isolated from marine environments (newly isolated strains group) and reference strains obtained from infected eels (reference strains group). The two groups had distinctly different profiles of the amplicons produced from RAPD. Additionally, biochemical comparison of these strains revealed that all four strains isolated from marine environments differed from the strains isolated from eels in their ability to promote D-mannitol fermentation. Two (NH 1 and NH 2) out of four isolates of biotype 2 from marine environments showed pathogenicity in eels Anguilla japonica in a challenge test. These isolates did not agglutinate with antisera against V. vulnificus NCIMB 2137 (serovar E), ATCC 27562 (non-serovar E), and ATCC 33816 (atypical serovar E).