• Title/Summary/Keyword: Subtyping

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Pulsed Field Gel Electrophoresis for Subtyping of Listeria monocytogenes

  • Jang, Sung-Sik;Fleet, Graham H.;Cox, Julian M.
    • Journal of Applied Biological Chemistry
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    • v.48 no.2
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    • pp.58-64
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    • 2005
  • Listeria monocytogenes is a high-risk foodborne pathogen responsible for foodborne listeriosis outbreaks, and is particularly dangerous to immuno-compromised people with mortality rate of about 30%. This review summarizes subtyping of L. monocytogenes using Pulsed Field Gel Electrophoresis, widely used to trace origin of foodborne outbreaks and to determine relationship between isolates.

PCR-RFLP for Rapid Subtyping of Plasmodium vivax Korean Isolates

  • Kang, Jung-Mi;Lee, Jinyoung;Kim, Tae Im;Koh, Eun-Ha;Kim, Tong-Soo;Sohn, Woon-Mok;Na, Byoung-Kuk
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.159-165
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    • 2017
  • Vivax malaria reemerged in Korea in 1993 and the outbreak has been continued with fluctuating numbers of annual indigenous cases. Understanding the nature of the genetic population of Plasmodium vivax circulating in Korea is beneficial for the knowledge of the nationwide parasite heterogeneity and in the implementation of malaria control programs in the country. Previously, we analyzed polymorphic nature of merozoite surface protein-1 (MSP-1) and MSP-$3{\alpha}$ in Korean P. vivax population and identified the Korean P. vivax population has been diversifying rapidly, with the appearance of parasites with new genetic subtypes, despite the recent reduction of the disease incidence. In the present study, we developed simple PCR-RFLP methods for rapid subtyping of MSP-1 and MSP-$3{\alpha}$ of Korean P. vivax isolates. These PCR-RFLP methods were able to easily distinguish each subtype of Korean P. vivax MSP-1 and MSP-$3{\alpha}$ with high accuracy. The PCR-RFLP subtyping methods developed here would be easily applied to massive epidemiological studies for molecular surveillance to understand genetic population of P. vivax and to supervise the genetic variation of the parasite circulating in Korea.

Development of Multiplex RT-PCR Assays for Rapid Detection and Subtyping of Influenza Type A Viruses from Clinical Specimens

  • Chang, Hee-Kyoung;Park, Jeung-Hyun;Song, Min-Suk;Oh, Taek-Kyu;Kim, Seok-Young;Kim, Chul-Jung;Kim, Hyung-Gee;Sung, Moon-Hee;Han, Heon-Seok;Hahn, Youn-Soo;Choi, Young-Ki
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1164-1169
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    • 2008
  • We developed multiplex RT-PCR assays that can detect and identify 12 hemagglutinin (H1-H12) and 9 neuraminidase (N1-N9) subtypes that are commonly isolated from avian, swine, and human influenza A viruses. RT-PCR products with unique sizes characteristic of each subtype were amplified by multiplex RT-PCRs, and sequence analysis of each amplicon was demonstrated to be specific for each subtype with 24 reference viruses. The specificity was demonstrated further with DNA or cDNA templates from 7 viruses, 5 bacteria, and 50 influenza A virus-negative specimens. Furthermore, the assays could detect and subtype up to $10^5$ dilution of each of the reference viruses that had an original infectivity titer of $10^6\;EID_{50}/ml$. Of 188 virus isolates, the multiplex RT-PCR results agreed completely with individual RT-PCR subtyping results and with results obtained from virus isolations. Furthermore, the multiplex RT-PCR methods efficiently detected mixed infections with at least two different subtypes of influenza viruses in one host. Therefore, these methods could facilitate rapid and accurate subtyping of influenza A viruses directly from field specimens.

PAIVS: prediction of avian influenza virus subtype

  • Park, Hyeon-Chun;Shin, Juyoun;Cho, Sung-Min;Kang, Shinseok;Chung, Yeun-Jun;Jung, Seung-Hyun
    • Genomics & Informatics
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    • v.18 no.1
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    • pp.5.1-5.5
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    • 2020
  • Highly pathogenic avian influenza (HPAI) viruses have caused severe respiratory disease and death in poultry and human beings. Although most of the avian influenza viruses (AIVs) are of low pathogenicity and cause mild infections in birds, some subtypes including hemagglutinin H5 and H7 subtype cause HPAI. Therefore, sensitive and accurate subtyping of AIV is important to prepare and prevent for the spread of HPAI. Next-generation sequencing (NGS) can analyze the full-length sequence information of entire AIV genome at once, so this technology is becoming a more common in detecting AIVs and predicting subtypes. However, an analysis pipeline of NGS-based AIV sequencing data, including AIV subtyping, has not yet been established. Here, in order to support the pre-processing of NGS data and its interpretation, we developed a user-friendly tool, named prediction of avian influenza virus subtype (PAIVS). PAIVS has multiple functions that support the pre-processing of NGS data, reference-guided AIV subtyping, de novo assembly, variant calling and identifying the closest full-length sequences by BLAST, and provide the graphical summary to the end users.

Molecular Typing in Public Health Laboratories: From an Academic Indulgence to an Infection Control Imperative

  • Allerberger, Franz
    • Journal of Preventive Medicine and Public Health
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    • v.45 no.1
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    • pp.1-7
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    • 2012
  • Using three Austrian case studies, the variegated applications of molecular typing in today's public health laboratories are discussed to help illustrate preventive management strategies relying on DNA subtyping. DNA macrorestriction analysis by pulsed field gel electrophoresis has become the gold standard for subtyping of food borne pathogens like listeria, salmonella, campylobacter and Bacillus cereus. Using a Salmonella Mbandaka outbreak from the year 2010 as example, it is shown how the comparison of patterns from human isolates, food isolates, animal isolates and feed isolates can allow to identify and confirm a source of disease. An epidemiological connection between the simultaneous occurrence of tuberculosis in cattle and deer with cases of human tuberculosis due to Mycobacterium caprae in 2010 was excluded using mycobacterial interspersed repetitive units variable-number tandem repeats subtyping. Also in 2010, multilocus sequence typing with nonselective housekeeping genes, the so-called sequence based typing protocol, was used to elucidate connections between an environmental source (a hospital drinking water system) and a case of legionellosis. During the last decades, molecular typing has evolved to become a routine tool in the daily work of public health laboratories. The challenge is now no longer to simply type microorganisms, but to type them in a way that allows for data exchange between public health laboratories all over the world.

Biopsy and Mutation Detection Strategies in Non-Small Cell Lung Cancer

  • Jung, Chi Young
    • Tuberculosis and Respiratory Diseases
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    • v.75 no.5
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    • pp.181-187
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    • 2013
  • The emergence of new therapeutic agents for non-small cell lung cancer (NSCLC) implies that histologic subtyping and molecular predictive testing are now essential for therapeutic decisions. Histologic subtype predicts the efficacy and toxicity of some treatment agents, as do genetic alterations, which can be important predictive factors in treatment selection. Molecular markers, such as epidermal growth factor receptor (EGFR) mutation and anaplastic lymphoma kinase (ALK) rearrangement, are the best predictors of response to specific tyrosine kinase inhibitor treatment agents. As the majority of patients with NSCLC present with unresectable disease, it is therefore crucial to optimize the use of tissue samples for diagnostic and predictive examinations, particularly for small biopsy and cytology specimens. Therefore, each institution needs to develop a diagnostic approach requiring close communication between the pulmonologist, radiologist, pathologist, and oncologist in order to preserve sufficient biopsy materials for molecular analysis as well as to ensure rapid diagnosis. Currently, personalized medicine in NSCLC is based on the histologic subtype and molecular status. This review summarizes strategies for tissue acquisition, histologic subtyping and molecular analysis for predictive testing in NSCLC.

Identification of ERBB pathway-activated cells in triple-negative breast cancer

  • Cho, Soo Young
    • Genomics & Informatics
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    • v.17 no.1
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    • pp.3.1-3.4
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    • 2019
  • Intratumor heterogeneity within a single tumor mass is one of the hallmarks of malignancy and has been reported in various tumor types. The molecular characterization of intratumor heterogeneity in breast cancer is a significant challenge for effective treatment. Using single-cell RNA sequencing (RNA-seq) data from a public resource, an ERBB pathway activated triple-negative cell population was identified. The differential expression of three subtyping marker genes (ERBB2, ESR1, and PGR) was not changed in the bulk RNA-seq data, but the single-cell transcriptomes showed intratumor heterogeneity. This result shows that ERBB signaling is activated using an indirect route and that the molecular subtype is changed on a single-cell level. Our data propose a different view on breast cancer subtypes, clarifying much confusion in this field and contributing to precision medicine.

Applications of MALDI-TOF Mass Spectrometry in Clinical Microbiology

  • Shin, Kyeong Seob;Yum, Jonghwa
    • Biomedical Science Letters
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    • v.28 no.3
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    • pp.145-156
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    • 2022
  • Over the past few decades, few technologies have had a greater impact on clinical microbiology laboratories than matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS). The MALDI-TOF MS is a fast, accurate, and low-cost and efficient method of microbial identification. This technology generates characteristic mass spectral fingerprints that is a unique signature for each microorganism, making it an ideal method for accurate identification at the genus and species levels of both bacterial and fastidious microorganism such as anaerobes, mycobacterium and fungi etc. In addition, MALDI-TOF MS has been successfully used in microbial subtyping and susceptibility tests such as determination of resistance genes. In this study, the authors summarized the application of MALDI-TOF MS in clinical microbiology and clinical research and explored the future of MALDI-TOF MS.

Classification of Listeria monocytogenes Isolates from Korean Domestic Foods Using Random Amplification of Polymorphic DNA and Serotyping Analysis (Random Amplification of Polymorphic DNA와 혈청학적 분석을 이용한 국내식품에서 분리한 Listeria monocytogenes의 분류)

  • Kim Hyun-Joong;Park Si-Hong;Kim Hae-Yeong
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.23-27
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    • 2006
  • Molecular subtyping of Listeria monocytogenes, including type strains and isolates from Korean foods, were performed using random amplification of polymorphic DNA (RAPD). Each Listeria species showed specific RAPD band patterns, and L. monocytogenes serotypes and isolates were divided into two clusters. RAPD results showed that L. monocytogenes isolates from Korean foods were divided into two groups. Group I contained L. monocytogenes serotypes 1/2b and 4b, whereas Group II contained serotypes 1/2a and 1/2c. These results suggested RAPD as possible subtyping methods for Listeria species. Also, RAPD Results showed significant correlation between molecular subtyping and serotyping of L. monocytogenes, and classified two different groups of L. monocytogenes isolated from Korean foods.