• Title/Summary/Keyword: Multiplex RT-PCR

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Detection and clinical manifestations of twelve respiratory viruses in hospitalized children with acute lower respiratory tract infections : Focus on human metapneumovirus, human rhinovirus and human coronavirus (하기도 감염으로 입원한 소아에서 12종 바이러스의 검출 및 임상 양상)

  • Kim, Kum Hyang;Lee, Jung Ho;Sun, Dong Shin;Kim, Yong Bae;Choi, Young Jin;Park, Joon Soo;Kim, Chang Jin;Jung, Dong Jun
    • Clinical and Experimental Pediatrics
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    • v.51 no.8
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    • pp.834-841
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    • 2008
  • Purpose : This study was perfomed to analyze in detail the viral etiology of acute lower respiratory tract infections (ALRI) in Cheunan, Korea by multiplex RT-PCR, including human rhinovirus (hRV) and newly identified viruses such as human metapneumovirus (hMPV) and human coronavirus (HCoV-OC43, HCoV-229E/NL63). Method : Nasopharyngeal aspirates (NPA) were collected from 863 hospitalized children with ALRI on the first day of admission at Soonchunhyang University Cheonan Hospital and analyzed by multiplex RT-PCR from December 2005 to November 2006. Results : Viral agents were detected from 474 subjects (54.9%). The identified viral pathogens were hRV 9.2%, hMPV 6.8%, HCoV-229E/NL63 1.4%, and HCoV-OC43 2.1%. Coinfections with ${\geq}2$ viruses were observed in 108 patients (22.8%). The major period of viral ALRI was the first year of life. Clinical diagnoses of viral ALRI were pneumonia (59.5%), bronchiolitis (24.7%), tracheobronchitis (11.4%), and croup (4%). The most common causes of bronchiolitis was respiratory syncytial virus B (RSV B), whereas hMPV, hRV, HCoV-229E/NL63, and HCoV-OC43 were commonly found in patients with pneumonia. The number of hMPV infections peaked between March and May 2006. HCoV-OC43 was prevalent from November to February 2006, whereas HCoV-229E and hRV were detected throughout the year. Conclusion : Although the study was confined to one year, hMPV was not detected during winter and peaked between March and April, which was not consistent with previous studies'. This present study indicates that HCoV is a less common respiratory pathogen in cases of ALRI in Korean children

Characteristics and Outcomes of Patients with Pulmonary Acute Respiratory Distress Syndrome Infected with Influenza versus Other Respiratory Viruses

  • Yoo, Jung-Wan;Ju, Sunmi;Lee, Seung Jun;Cho, Min-Chul;Cho, Yu Ji;Jeong, Yi Yeong;Lee, Jong Deog;Kim, Ho Choel
    • Tuberculosis and Respiratory Diseases
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    • v.82 no.4
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    • pp.328-334
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    • 2019
  • Background: Although the frequency of respiratory viral infection in patients with pulmonary acute respiratory distress syndrome (ARDS) is not uncommon, clinical significance of the condition remains to be further elucidated. The purpose of this study was to compare characteristics and outcomes of patients with pulmonary ARDS infected with influenza and other respiratory viruses. Methods: Clinical data of patients with pulmonary ARDS infected with respiratory viruses January 2014-June 2018 were reviewed. Respiratory viral infection was identified by multiplex reverse transcription-polymerase chain reaction (RT-PCR). Results: Among 126 patients who underwent multiplex RT-PCR, respiratory viral infection was identified in 46% (58/126): 28 patients with influenza and 30 patients with other respiratory viruses. There was no significant difference in baseline and clinical characteristics between patients with influenza and those with other respiratory viruses. The use of extracorporeal membrane oxygenation (ECMO) was more frequent in patients with influenza than in those with other respiratory viruses (32.1% vs 3.3%, p=0.006). Co-bacterial pathogens were more frequently isolated from respiratory samples of patients with pulmonary ARDS infected with influenza virus than those with other respiratory viruses. (53.6% vs 26.7%, p=0.036). There were no significant differences regarding clinical outcomes. In multivariate analysis, acute physiology and chronic health evaluation II was associated with 30-mortality (odds ratio, 1.158; 95% confidence interval, 1.022-1.312; p=0.022). Conclusion: Respiratory viral infection was not uncommon in patients with pulmonary ARDS. Influenza virus was most commonly identified and was associated with more co-bacterial infection and ECMO therapy.

Genetic Diversity of Foodborne Pathogen Detected in Commercial Shellfish in Metropolitan Area

  • Park, Jung Hun;Cho, Kyu Bong
    • Biomedical Science Letters
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    • v.28 no.2
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    • pp.83-91
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    • 2022
  • This study was performed to investigate the contamination status of microorganisms that cause food poisoning in shellfish distributed in the metropolitan area of South Korea. The analyses were conducted according to the sample type, season, and region. In particular, the genotype of food poisoning viruses that explosively cause collective food poisoning was analyzed. Total 483 each of four types of shellfish (Crassostrea gigas, Mytilus coruscus, Pectea albicans albicans, and Scapharca subcrenata) were collected from four distribution sites located in the metropolitan area. We investigated foodborne pathogen by multiplex PCR and RT/Nested PCR from shellfish. As a result, Vibrio parahaemolyticus and Bacillus cereus were detected in 13 and 21 samples and Norovirus (NoV) GI and GII were detected in 7 and 12 samples, respectively. V. parahaemolyticus and NoV GI and GII showed differences in types of shellfish and seasons, and no correlation was confirmed with regional differences. Also, as a result of genotyping for the detected NoV GI and GII, they were confirmed as NoV GI.7, GI.5 and GII.3, which are causes of collective food poisoning. Therefore, cross-infection with shellfish can cause food poisoning. In particular, attention must be paid to the handling and cooking of shellfish in organizations that implement group feeding, and it is necessary to establish a management system for microorganisms that cause food poisoning in the process of shellfish farming and distribution.

Clinical characteristics of acute lower respiratory tract infections due to 13 respiratory viruses detected by multiplex PCR in children (소아에서 13종 호흡기 바이러스에 의한 급성 하기도 감염의 임상 양상)

  • Lim, Jeong-Sook;Woo, Sung-Il;Baek, Yun-Hee;Kwon, Hyuk-Il;Choi, Young-Ki;Hahn, Youn-Soo
    • Clinical and Experimental Pediatrics
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    • v.53 no.3
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    • pp.373-379
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    • 2010
  • Purpose : This study was performed to investigate the epidemiologic and clinical features of 13 respiratory viruses in children with acute lower respiratory tract infections (ALRIs). Methods : Nasopharyngeal aspirates were prospectively obtained from 325 children aged 15 years or less from May 2008 to April 2009 and were tested for the presence of 13 respiratory viruses by multiplex real-time-polymerase chain reaction (RT-PCR). Results : Viruses were identified in 270 children (83.1%). Co-infections with ${\geq}2$ viruses were observed in 71 patients (26.3 %). Respiratory syncytial virus (RSV) was the most common virus detected (33.2%), followed by human rhinovirus (hRV) (19.1%), influenza virus (Flu A) (16.9%), human metapneumovirus (hMPV) (15.4%), parainfluenza viruses (PIVs) (8.3%), human bocavirus (hBoV) (8.0%), adenovirus (ADV) (5.8%), and human coronavirus (hCoV) (2.2%). Clinical diagnoses of viral ALRIs were bronchiolitis (37.5%), pneumonia (34.5%), asthma exacerbation (20.9%), and croup (7.1%). Clinical diagnoses of viral bronchiolitis and pneumonia were frequently demonstrated in patients who tested positive for RSV, hRV, hMPV, or Flu A. Flu A and hRV were most commonly identified in children older than 3 years and were the 2 leading causes of asthma exacerbation. hRV C was detected in 14 (4.3%) children, who were significantly older than those infected with hRV A ($mean{\pm}SD$, $4.1{\pm}3.5$ years vs. $1.7{\pm}2.3$ years; P =0.009). hBoV was usually detected in young children ($2.3{\pm}3.4$ years) with bronchiolitis and pneumonia. Conclusion : This study described the features of ALRI associated with 13 respiratory viruses in Korean children. Additional investigations are required to define the roles of newly identified viruses in children with ALRIs.

Development of Diagnostic System to Black Queen Cell Virus(BQCV) Using Multi-point Detection (Multi-point PCR법을 이용한 Black Queen Cell Virus (BQCV) 검출법 개발)

  • Kim, Somin;Kim, Byounghee;Kim, Moonjung;Kim, Jungmin;Truong, A Tai;Kim, Seonmi;Yoon, Byoungsu
    • Journal of Apiculture
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    • v.34 no.1
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    • pp.39-46
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    • 2019
  • BQCV multi-point PCR was developed as a rapid multiplex detection method for BQCV, one of the viral pathogens of honeybees. It could detect BQCV specific genes qualitative as well as quantitative detection based on ultra-rapid PCR. Three primer pairs (RNA dependent RNA polymerase, capsid protein, 3C like protease) were specifically designed for accurate the detection and were optimized for minimizing the detection time and increasing the sensitivity. Our advanced diagnostic system have the accuracy by lowering the concern about the variation in the BQCV detection site. In addition, it should be an opportunity to identify mutations that are mixed with other viruses.

Active Surveillance of Pertussis in Infants Under 6 Months of Age: A Single Center Experience from 2011 to 2013 (생후 6개월 미만의 영아에서 백일해에 대한 적극적인 감시활동: 2011-2013년 단일기관 연구)

  • Han, Young Ik;Choi, Ji Yeon;Lee, Hyewon;Lee, Teak-Jin
    • Pediatric Infection and Vaccine
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    • v.21 no.2
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    • pp.114-120
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    • 2014
  • Purpose: The objective of this study was to assess and investigate the epidemiology of pertussis in infants under 6 months of age. Methods: A prospective study was conducted between October 1, 2011 and April 30, 2013 in CHA Bundang Medical Center, Seongnam, South Korea. Polymerase chain reaction (PCR) or culture was used to detect Bordetella pertussis in nasopharyngeal aspirates from case patients who were hospitalized for acute lower respiratory tract infection (LRTI). In addition, multiplex real-time PCR assays were also performed to detect 6 etiologic viruses, including adenovirus, human metapeumo-virus, influenza virus, parainfluenza virus, respiratory syncytial virus and rhinovirus. Results: Of the 79 enrolled case patients, whose median age was 2 months of age, the most common diagnoses uncovered in this study were acute bronchiolitis (60%) and pneumonia (28%). B. pertussis infection was found in 13 cases (16%), in which 7 (53%) was coinfected with respiratory syncytial virus and 1 (7%) with influenza A virus. Of the 13 patients with B. pertussis infection, 6 (46%) were not vaccinated with the diphtheria, tetanus toxoid, and acellular pertussis vaccine, while 6 (46%) received 1 dose, and 1 (8%) received 2 doses. Conclusion: B. pertussis infection was present in 16% of under 6 month-old infants, who were hospitalized for acute LRTI. Therefore, a nationwide epidemiological surveillance of pertussis, including institutions that cater to infants under 6 months of age is necessary and needed.

Typing and Sequence Analysis of the VP7 Gene of Rotavirus Isolated from Infants in Korea (한국영아에서 분리된 로타바이러스의 VP7 유전자형 및 염기서열 분석)

  • Song, Mi-Ok;Yun, Karyn;Chung, Sang-In;Choi, Chul-Soon;Lim, In-Seok;Kang, Shien-Young;An, Chang-Nam;Kim, Won-Yong
    • The Journal of Korean Society of Virology
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    • v.30 no.2
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    • pp.101-112
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    • 2000
  • Rotaviruses are the most common cause of severe vomiting and diarrhea in children worldwide and classified as a genus in the family Reoviridae. Rotavirus has eleven segmented dsRNAs and the virion consists of three shells. Outer capsid VP7 and VP4 induce neutralizing antibodies and are classified into G types (glycoprotein VP7) and P types (protease-sensitive VP4). Characterization of VP7 gene of Korean isolates of human rotavirus was performed using multiplex PCR and nucleotide sequence analysis. After RT-PCR amplification of full length (1,062 bp) of VP7 genes, the amplified PCR products were G typed by multiplex PCR and the nucleotide sequences were compared with those of reference rotavirus from GenBank. The G type analysis revealed that 25% (2/8) belong to G1, whereas 37.5% (3/8) benong to G2 and G4, respectively. The Korean isolates within the same serotypes showed high homology of nucleotide sequences and could be discriminated from foreign isolates exception with two strains (CAU009 and CAU022). But Korean isolates CAU009 and CAU022 were close related into japanease isolates 417 (99.2%) and indian isolates (97.6%) than Korean isolatese. Our results showed that these two strains were supposed to be originated from abroad. As a results, The G typing and nucleotide sequence analysis of VP7 gene of rotavirus isolated from infants in Korea could be used for identification, serotying and determination of novel or unusual strains of rotaviruses.

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Acute viral gastroenteritis in children hospitalized in Iksan, Korea during December 2010-June 2011

  • So, Cheol Whoan;Kim, Dong Sup;Yu, Seung Taek;Cho, Ji-Hyun;Kim, Jong Duck
    • Clinical and Experimental Pediatrics
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    • v.56 no.9
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    • pp.383-388
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    • 2013
  • Purpose: Viral etiology is common in cases of children with acute diarrhea, and antibiotic therapy is usually not required. Therefore, it is important to determine the distribution of common viruses among children hospitalized with acute diarrhea. Methods: We included 186 children who suffered from acute diarrhea and were hospitalized at the Wonkwang University Hospital Pediatric ward from December 1, 2010 to June 30, 2011 in this study. Stool samples were collected and multiplex reverse transcriptase polymerase chain reaction (multiplex RT-PCR) was used to simultaneously determine the viral etiology such as rotavirus, norovirus, astrovirus, or adenovirus. Results: Causative viruses were detected in 72 of the 186 cases (38.7%). The mean age of the virus-positive cases was 1 year and 9 months (range, 1 month to 11 years). Rotavirus was detected in 50/186 (26.9%); norovirus, in 18/186 (9.7%); and astrovirus, in 3/186 cases (1.6%). Adenovirus was not detected in any of the cases. Proportions of norovirus genogroups I and II were 21.1% and 78.9%, respectively. Four of the 51 rotavirus-positive cases (7.8%) had received rotavirus vaccination at least once. The mean duration of diarrhea was 2.8 days (range, 1 to 10 days) and vomiting occurred in 39 of the 72 cases (54.2%). Conclusion: Viral etiology was confirmed in about one-third of the children with acute diarrhea, and the most common viral agent was rotavirus, followed by norovirus.

Clinical and laboratory profiles of hospitalized children with acute respiratory virus infection

  • Choi, Eunjin;Ha, Kee-Soo;Song, Dae Jin;Lee, Jung Hwa;Lee, Kwang Chul
    • Clinical and Experimental Pediatrics
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    • v.61 no.6
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    • pp.180-186
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    • 2018
  • Purpose: Despite the availability of molecular methods, identification of the causative virus in children with acute respiratory infections (ARIs) has proven difficult as the same viruses are often detected in asymptomatic children. Methods: Multiplex reverse transcription polymerase chain reaction assays were performed to detect 15 common respiratory viruses in children under 15 years of age who were hospitalized with ARI between January 2013 and December 2015. Viral epidemiology and clinical profiles of single virus infections were evaluated. Results: Of 3,505 patients, viruses were identified in 2,424 (69.1%), with the assay revealing a single virus in 1,747 cases (49.8%). While major pathogens in single virus-positive cases differed according to age, human rhinovirus (hRV) was common in patients of all ages. Respiratory syncytial virus (RSV), influenza virus (IF), and human metapneumovirus (hMPV) were found to be seasonal pathogens, appearing from fall through winter and spring, whereas hRV and adenovirus (AdV) were detected in every season. Patients with ARIs caused by RSV and hRV were frequently afebrile and more commonly had wheezing compared with patients with other viral ARIs. Neutrophil-dominant inflammation was observed in ARIs caused by IF, AdV, and hRV, whereas lymphocyte-dominant inflammation was observed with RSV A, parainfluenza virus, and hMPV. Monocytosis was common with RSV and AdV, whereas eosinophilia was observed with hRV. Conclusion: In combination with viral identification, recognition of virus-specific clinical and laboratory patterns will expand our understanding of the epidemiology of viral ARIs and help us to establish more efficient therapeutic and preventive strategies.

Identification of Viral Pathogens for Lower Respiratory Tract Infection in Children at Seoul During Autumn and Winter Seasons of the Year of 2008-2009 (서울의 2008-2009년 가을, 겨울철에 유행한 소아의 하기도 호흡기 바이러스 병원체의 규명)

  • Kim, Ki Hwan;Kim, Ji Hong;Kim, Kyung Hyo;Kang, Chun;Kim, Ki Soon;Chung, Hyang Min;Kim, Dong Soo
    • Pediatric Infection and Vaccine
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    • v.17 no.1
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    • pp.49-55
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    • 2010
  • Purpose : The Purposes of this study are to identify the circulating etiologic viruses of acute lower respiratory tract infection in children and to understand the relation with clinical diagnosis. Methods : We obtained a total of 418 nasopharyngeal aspirates from children admitted for their acute lower respiratory tract infections at three tertiary hospitals in Seoul from September 2008 to March 2009. We performed multiplex RT-PCR to identify 14 etiologic viruses and analyzed their emerging patterns and clinical features. Results : Average age of patients was 16.4 months old and the ratio of male to female was 1.36. Viruses were detected in 56.2% of a total of 418 samples. Respiratory syncytial virus (35%) was the most frequently detected and followed by human rhinovirus (22%), human bocavirus, adenovirus, human metapneumovirus, parainfluenza virus, influenza virus and human coronavirus. Co-infection reached 21.9 % of positive patients. Conclusion : When we manage the patients with acute lower respiratory infectious diseases, we should remind the role of various viral pathogens, which might be circulating by seasons and by local areas.