• Title/Summary/Keyword: Multiplex real-time PCR

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Supraclavicular BCG Lymphadenitis Noted at 21 Months after BCG Vaccination Confirmed by a Molecular Method (분자생물학적 방법으로 확진한 접종 21개월 후에 발생한 BCG 쇄골상부 림프절염 1례)

  • Lee, Min Hyun;Chae, Moon-Hee;Park, Kyoung Un;Cho, Hye-Kyung
    • Pediatric Infection and Vaccine
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    • v.21 no.2
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    • pp.139-143
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    • 2014
  • Bacille Calmette-$Gu\acute{e}rin$ (BCG) lymphadenitis is the most common complication of BCG vaccination. It commonly occurs in infants aged <6 months involving ipsilateral axillary lymph nodes. We described BCG lymphadenitis in a 22-month-old boy presenting swelling of left supraclavicular lymph node that was confirmed by real-time polymerase chain reaction (PCR) and the multiplex PCR targeting the region of difference (RD).

High Prevalence of Mycobacterium avium subsp. paratuberculosis in Wild Ducks in the Middle Area of South Korea

  • Rhim, Haerin;Cho, Yong-Il;Jang, Hye-Jin;Na, Ki-Jeong;Han, Jae-Ik
    • Journal of Veterinary Clinics
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    • v.35 no.1
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    • pp.7-9
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    • 2018
  • Mycobacterium avium subspecies paratuberculosis (MAP) causes a significant economic burden in the animal production industry. The purpose of this study was to investigate the prevalence of MAP in the feces of wild duck populations residing along a riverside close to farms in the center of Korea. From wild Spot-billed (Anas poecilorhyncha) and Mallard (Anas platyrhynchos) ducks, 128 fecal samples were collected and analyzed using multiplex real-time PCR, sequencing, and nested PCR to confirm the presence of the organism. The molecular analyses showed that 44 samples (34.4%) were positive for MAP, suggesting a high prevalence of MAP in the wild duck population. Considering the nature and habitat of wild ducks, this result suggests that the organism was introduced from contaminated water from waste of nearby farms, and that the wild ducks may act as a transmitter of the organism to other wild birds or livestock.

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.

The Uneven Distribution of Mating Type Genes in Natural and Cultivated Truffle Orchards Contributes to the Fructification of Tuber indicum

  • Li, Qiang;Fu, Yu;Sun, Qun;Sourzat, Pierre;Yang, Mei;Liu, Chengyi;Tan, Hao;Ye, Lei;Zou, Jie;Wu, Chenguang;Zhang, Bo;Li, Xiaolin
    • Mycobiology
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    • v.46 no.1
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    • pp.64-71
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    • 2018
  • The aim of this study was to investigate the pattern of distribution of mating type (MAT) genes of Tuber indicum in ectomycorhizosphere soils from natural T. indicum-producing areas and cultivated truffle orchards and ascocarp samples from different regions. Quantitative real-time PCR and multiplex PCR were used to weight the copy numbers of MAT1-1-1 and MAT1-2-1 in natural truffle soils and cultivated orchard soils. The effect of limestone on the pattern of truffle MAT genes and the correlation between soil properties and the proportion of MAT genes were also assessed. These results indicated that an uneven and nonrandom distribution of MAT genes was common in truffle-producing areas, cultivated truffle orchards, and ascocarps gleba. The competition between the two mating type genes and the expansion of unbalanced distribution was found to be closely related to truffle fructification. Limestone treatments failed to alter the proportion of the two mating type genes in the soil. The content of available phosphorus in soil was significantly correlated with the value of MAT1-1-1/MAT1-2-1 in cultivated and natural ectomycorhizosphere soils. The application of real-time quantitative PCR can provide reference for monitoring the dynamic changes of mating type genes in soil. This study investigates the distributional pattern of T. indicum MAT genes in the ectomycorhizosphere soil and ascocarp gleba from different regions, which may provide a foundation for the cultivation of T. indicum.

Microbial Community Analysis of a Methane-Oxidizing Biofilm Using Ribosomal Tag Pyrosequencing

  • Kim, Tae-Gwan;Lee, Eun-Hee;Cho, Kyung-Suk
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.360-370
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    • 2012
  • Current ecological knowledge of methanotrophic biofilms is incomplete, although they have been broadly studied in biotechnological processes. Four individual DNA samples were prepared from a methanotrophic biofilm, and a multiplex 16S rDNA pyrosequencing was performed. A complete library (before being de-multiplexed) contained 33,639 sequences (average length, 415 nt). Interestingly, methanotrophs were not dominant, only making up 23% of the community. Methylosinus, Methylomonas, and Methylosarcina were the dominant methanotrophs. Type II methanotrophs were more abundant than type I (56 vs. 44%), but less richer and diverse. Dominant non-methanotrophic genera included Hydrogenophaga, Flavobacterium, and Hyphomicrobium. The library was de-multiplexed into four libraries, with different sequencing efforts (3,915 - 20,133 sequences). Sorrenson abundance similarity results showed that the four libraries were almost identical (indices > 0.97), and phylogenetic comparisons using UniFrac test and P-test revealed the same results. It was demonstrated that the pyrosequencing was highly reproducible. These survey results can provide an insight into the management and/or manipulation of methanotrophic biofilms.

Carbapenemase-Producing Enterobacterales: Epidemiology, Detection, and Treatment

  • Yun Hee Baek;Kyeong Seob Shin
    • Biomedical Science Letters
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    • v.29 no.3
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    • pp.109-120
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    • 2023
  • Recently, the explosive increase of carbapenemase-producing Enterobacterales (CPE) in the worldwide poses a serious threat. The purpose of this study is to investigate epidemiology, detection, and treatment of CPE. Three main carbapenemase are reported worldwide, which were KPC, NDM, and OXA-48-like. KPC type are mostly found in USA, China, Europe, and Latin America. NDM type are mostly found in South Asia. OXA-48-like are often seen in the Mediterranean and Northern Africa. In Korea, CPE have increased explosively since 2015. In 2021, 18,099 CPE were isolated, which were Klebsiella pneumoniae, Escherichia coli, and Enterobacter cloacae in order. The CPE genotype was distributed with KPC, NDM, OXA type in order. Phenotypic detection methods include carbapenemase production tests (CPT) and differential tests of CPE. CPTs include modified Hodge test, modified carbapenem inactivation method (mCIM), Carba NP test, among which mCIM is the most widely used due to easy accessibility and accuracy. A lot of genotypic methods are being done for quick results, and commercialized kits using multiplex real-time PCR and microarray are widely used. Colistin and tigecycline are used as the first line of CPE treatment and are used in combination with second line drugs such as meropenem and fosfomycin.

Detection of Airborne Respiratory Viruses in Residential Environments (주거환경 공기 중 호흡기 바이러스의 검출)

  • Park, Keun-Tae;Moon, Kyong-Whan;Kim, Hyung-Tae;Park, Chan-Jung;Jeong, Ho-Chul;Lim, Young-Hee
    • Journal of Environmental Health Sciences
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    • v.37 no.4
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    • pp.306-314
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    • 2011
  • Objectives: Respiratory virus infections are the most common disease among all ages in all parts of the world and occur through airborne transmission. The purpose of this study was to detect and quantitate human respiratory viruses in residential environments. Methods: Air samples were collected from the residential space of apartments in the Seoul/Gyeonggi-do area. The samples were collected from indoor and outdoor air. Among respiratory viruses, influenza A virus, influenza B virus, parainfluenza virus, metapneumovirus, respiratory syncytial virus, and adenovirus were investigated by multiplex polymerase chain reaction. Among the virus-positive samples, we performed adenovirus quantification by real-time polymerase chain reaction. Results: Virus detection rates were 44.0%, 3.8%, 3.4%, and 17.3% in spring, summer, autumn, and winter, respectively. The virus detection rate was higher in winter and spring than in summer and autumn. Adenovirus was most commonly detected, followed by influenza A virus and parainfluenza virus. Virus distribution was not significantly different between indoor and outdoor environments. Conclusions: Although virus concentrations were not high in residential environments, residents in houses with detected viruses may have an increased risk of exposure to airborne respiratory viruses, especially in winter and spring.

Pyramiding transgenes for potato tuber moth resistance in potato

  • Meiyalaghan, Sathiyamoorthy;Pringle, Julie M.;Barrell, Philippa J.;Jacobs, Jeanne M.E.;Conner, Anthony J.
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.293-301
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    • 2010
  • The feasibility of two strategies for transgene pyramiding using Agrobacterium-mediated transformation was investigated to develop a transgenic potato (Solanum tuberosum L. cv. Iwa) with resistance to potato tuber moth (PTM) (Phthorimaea operculella (Zeller)). In the first approach, cry1Ac9 and cry9Aa2 genes were introduced simultaneously using a kanamycin (nptII) selectable marker gene. The second approach involved the sequential introduction (re-transformation) of a cry1Ac9 gene, using a hygromycin resistance (hpt) selectable marker gene, into an existing line transgenic for a cry9Aa2 gene and a kanamycin resistance (nptII) selectable marker gene. Multiplex polymerase chain reaction (PCR) confirmed the presence of the specific selectable marker gene and both cry genes in all regenerated lines. The relative steady-state level of the cry gene transcripts in leaves was quantified in all regenerated lines by real-time PCR analysis. Re-transformation proved to be a flexible approach to effectively pyramid genes for PTM resistance in potato, since it allowed the second gene to be added to a line that was previously identified as having a high level of resistance. Larval growth of PTM was significantly inhibited on excised greenhouse-grown leaves in all transgenic lines, although no lines expressing both cry genes exhibited any greater resistance to PTM larvae over that previously observed for the individual genes. It is anticipated that these lines will permit more durable resistance by delaying the opportunities for PTM adaptation to the individual cry genes.

Respiratory Virus Detection Rate in Patients with Severe or Atypical Community-acquired Pneumonia (중증 또는 비전형적 지역사회획득 폐렴으로 입원한 환자에서 호흡기 바이러스의 검출 빈도)

  • Park, Ji-Won;Jung, Sun-Young;Eun, Hyuk-Soo;Cheon, Shin-Hye;Seong, Seok-Woo;Park, Dong-Il;Park, Myung-Rin;Park, Hee-Sun;Jung, Sung-Soo;Kim, Ju-Ock;Kim, Sun-Young;Lee, Jeong-Eun
    • Tuberculosis and Respiratory Diseases
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    • v.71 no.5
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    • pp.335-340
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    • 2011
  • Background: Community-acquired pneumonia (CAP) is an important cause of morbidity and mortality throughout the world in all age groups. Viral causes of CAP are less well characterized than bacterial causes. We analyzed the characteristics of hospitalized patients with CAP who had a viral pathogen detected by multiplex polymerase chain reaction (PCR). Methods: Multiplex real-time PCR was performed for respiratory viruses in samples collected from 520 adults who developed CAP at Chungnam National University Hospital. Clinical, laboratory, and radiological features at presentation as well as other epidemiological data were analyzed. Results: Of 520 patients with CAP, a viral pathogen was detected in 60 (11.5%), and influenza A was the most common. The virus detection rate in patients with CAP was highest in November. Two or more pathogens were detected in 13 (21.7%) patients. Seven patients had severe disease and were administered in the intensive care unit. Most patients (49/60, 81.7%) had comorbidities. However, nine (15%) patients had no comorbidities, and their age was <60 years. The ground glass opacity pattern was the most common radiological feature. Seven (11.7%) patients died from CAP. Conclusion: Viral pathogens are commonly detected in patients with CAP, and a respiratory virus may be associated with the severity and outcome of pneumonia. Careful attention should be paid to the viral etiology in adult patients with CAP.

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.