• Title/Summary/Keyword: 중합연쇄반응

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Sex Determination used Sex Determining Region Y Gene on the Y-chromosome of Human Teeth (사람 치아 Y염색체상의 sex determining region Y(SRY)유전자를 이용한 성별감정)

  • Kim, Sei-Youn;Ahn, Jong-Mo;Ryu, Geun-Chun;Yoon, Chang-Lyuk
    • Journal of Oral Medicine and Pain
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    • v.24 no.3
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    • pp.325-333
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    • 1999
  • 최근 중합효소연쇄반응을 이용한 분자생물학적 유전자분석기술의 발달로 성염색체상의 유전좌위 증폭을 통한 성별감정이 활발히 이루어지고 있다. 그중 사람 Y염색체상에 존재하는 남성 고환의 형성을 유도하는 sex-determining region Y(SRY) gene이 규명되어 유전질환의 조기 발견이나 예방 및 태아의 성별판정 등에 응용되고 있다. 그러나, 치아는 외부 환경에 대한 저항성이 가장 높은 장기로 성별감정 등 법의치과학적 개인식별에 널리 이용되고 있음에도 불구하고, SRY 유전자를 이용하여 치아에서의 성별감정에 대한 연구는 시도된 바 없다. 따라서, 본 연구에서는 사람 치아에서 중합효소연쇄반응법을 이용한 SRY 유전자를 검출하여 성별판정에 용용하고자 하였다. 남녀 각각 20개 치아의 치수와 상아질에서 DNA를 추출하여 중합효소연쇄반응 을 시행하고 SRY 유전자를 검색한 결과, 남성에서는 치수 13개중 8개, 상아질 7개중 4개에서 SRY 유전자가 검출되었고, 여생에서는 검출되지 않았다. 이러한 결과는 중합효소연쇄반응법을 이용하여 사람 치아에서 SRY 유전자를 검색할 때, 남성판별에 유용하고 치아를 이용한 성별감정시 기존의 성별감정에 이용되고 있는 다른 유전자와 함께 SRY 유전자를 검색함으로써 성별감정의 신뢰도를 높힐 수 있을 것으로 사료된다.

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Development of a Panel of Multiplex Real-Time Polymerase Chain Reaction Assays for Simultaneous Detection of Canine Enteric Bacterial Pathogens (개의 장내 병원균의 동시 검출을 위한 다중 실시간 중합효소연쇄반응분석 패널개발)

  • Jang, Hye-Jin;Han, Jae-Ik;Kang, Hyo-Min;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.32 no.2
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    • pp.154-157
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    • 2015
  • A major cause of diarrhea in a dog is an infection with bacteria which include Salmonella spp., Campylobacter (C.) spp., and Clostridium (Cl.) spp.. It is fastidious to identify these bacteria by the culture. The purpose of this experiment is to devise the method for detecting Cl. perfringens, C. jejuni, C. coli, and Salmonella spp. with rapid and high sensitivity. The fecal samples collected from 71 normal and 66 diarrheic dog feces were used to compare the prevalence of the enteric pathogens and to develop a multiplex real-time polymerase chain reaction (PCR) assay for clinical use. Detection of Cl. perfringens, C. coli, and C. jejuni in diarrhea feces was higher than normal feces. A developed multiplex real-time PCR is useful for determining the presence and quantity of pathogen-specific or other unique sequences with in a fecal sample.

A Study on the Distribution of Mycoplasma pneumoniae Infectious Disease in Children from 2016 to 2018 According to Age (2016년에서 2018년의 소아 마이코플라즈마폐렴균 감염질환의 연령별 발생분포에 대한 연구)

  • Junhyung Choi;Ki Hwan Kim
    • Pediatric Infection and Vaccine
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    • v.30 no.3
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    • pp.121-128
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    • 2023
  • Purpose: This study aimed to investigate the recent age distribution of Mycoplasma pneumoniae in patients with respiratory infections and the differences in diagnostic usefulness according to the methods used in these patients. Methods: We retrospectively reviewed the medical records of patients aged ≤18 years with respiratory infectious diseases who underwent polymerase chain reaction (PCR) or a specific immunoglobulin M (IgM) test between July 2016 and February 2019. The diagnosis of M. pneumoniae infection was confirmed by a positive result in the PCR or IgM test. Results: Of the 2,721 patients tested for M. pneumoniae, 2,197 underwent IgM, and 1,144 underwent PCR, with positivity rates of 17% and 20%, respectively. Among the 620 patients tested for both IgM and PCR tests simultaneously, 35% had M. pneumoniae infection, with 14% under 1 year old and 13% under 1-2 years old. The positive rate increased with age in both tests. Higher positive rates were observed in the IgM test before 3 years of age and in the PCR test after 3 years of age. The agreement rate between the two tests was 77.9% (Cohen's kappa 0.402). Conclusions: As age increased, the rates of M. pneumoniae infection also increased. In patients under 2 years of age, 4¬-14% of infections were confirmed depending on the method used. The moderate agreement between the PCR and IgM tests suggests that the simultaneous use of PCR and the IgM test for the early diagnosis should be approached with caution.

PCR-Based Sensitive Detection and Identification of Xanthomonas oryzae pv. oryzae (중합효소연쇄 반응에 의한 벼 흰잎마름병균의 특이적 검출)

  • Lee, Byoung-Moo;Park, Young-Jin;Park, Dong-Suk;Kim, Jeong-Gu;Kang, Hee-Wan;Noh, Tae-Hwan;Lee, Gil-Bok;Ahn, Joung-Kuk
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.256-264
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    • 2004
  • A new primer set was developed for the detection and identification of Xanthomonas oryzae pv. oryzae, the bacterial leaf blight (BLB) pathogen in rice plant. The nucleotide sequence of hpaA gene was determined from X. o. pv. oryzae str. KACC10331, and the sequence information was used to design primers for the application of the polymerase chain reaction (PCR). The nucleotide sequence of hpaA from X. o. pv. oryzae str. KACC 10331 was aligned with those of X. campestris pv. vesicatoria, X. campestris pv. campestris, X. axonopodis pv. citri, and X. axonopodis pv. glycines. Based on these results, a primer set(XOF and XOR) was designed for the specific detection of hpaA in X. o. pv. oryzae. The length of PCR products amplified using the primer set was 534-bp. The PCR product was detected from only X. o. pv. oryzae among other Xanthomonas strains and reference bacteria. This product was used to confirm the conservation of hpaA among Xanthomonas strains by Southern-blotting. Furthermore, PCR amplification with XOF and XOR was used to detect the pathogen in an artificially infected leaf. The sensitivity of PCR detection in the pure culture suspension was also determined. This PCR-based detection methods will be a useful method for the detection and identification of X. o. pv. oryzae as well as disease forecasting.

Development of In situ PCR Method Using Primer Polymers (프라이머 중합체를 이용한 원위치 중합효소 연쇄반응 In situ PCR 방법의 개발)

  • 장진수;이재영
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.167-171
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    • 2004
  • Reduction in the leakage of the amplified PCR product out of cell is required for effective in situ PCR. For this purpose, primers with complementary tail sequences at their 5' sides were utilized to synthesize high molecular weight PCR products, but it is time-consuming and causes deterioration of cellular appearance with many PCR cycles. Therefore, it is required to optimize the PCR condition with minimal PCR cycles. To achieve the pur-pose, primer polymers were made without the target DNA in tube from nonspecific amplification with tailed primers and treated onto the fixed Molt/LAV cells on the glass slide for the 20 cycle-in situ PCR, in which the appropriate target signals were observed for the possible use of primer polymers in in situ PCR.

Controlled Radical Polymerization of Vinyl Acetate in the presence of Tridecafluoro-n-hexyliodide(III) (비닐아세테이트의 조절된 라디칼 중합. III (요도드화물 존재 하에서))

  • 마석일;한규찬;김용일;권순홍
    • Proceedings of the Korean Fiber Society Conference
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    • 2002.04a
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    • pp.25-28
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    • 2002
  • 비공역형 단량체인 비닐아세테이트(VAc)는 공역형 비닐계 단량체와 달리 라디칼의 활성이 너무 커서 중합 도중 빈번한 연쇄이동반응과 정지반응에 의해 분지구조의 고분자가 얻어지며 고분자량의 폴리비닐아세테이트를 얻기가 어려운 것으로 알려져 있다. 폴리비닐알코올(PVA)은 비닐알코을 단량체의 호변이성질화 때문에 단량체의 직적중합에 의해서는 얻을 수 없고 일반적으로 비닐아세테이트(VAc)를 라디칼 중합하여 얻어진 폴리비닐아세테이트(PVAc)를 비누화하여 합성한다. (중략)

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Application of Digital Polymerase Chain Reaction for Human Gene Detection in Heterologous Tissues (이종 조직에서 사람 유전자 검출을 위한 디지털 중합효소연쇄반응의 적용)

  • Kim, Jin-Hee
    • Proceedings of the Korea Contents Association Conference
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    • 2018.05a
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    • pp.561-562
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    • 2018
  • 디지털 중합효소연쇄반응(Digital PCR)은 3세대 PCR로 명명하며, 1세대인 일반 PCR과 2세대인 정량 PCR(Real-time PCR)의 단점을 보완하여 개발된 방법이다. Digital PCR System은 소량의 PCR 반응을 10만개 이상의 반응통(wall)에 적재하는 방식의 나노유체칩에서 쪼개어 증폭시킨 후, target DNA를 계수한다. Target DNA의 증폭 여부에 따라 positive(1)와 negative(0)로 digital signal처럼 받아들여 계수하고, 포아송 분포를 통해 target DNA의 copy를 계산해 최종적으로 샘플 microlitr당 Copy수로 결과 값을 확인할 수 있다. 본 연구에서는 종(種)이 다른 동물의 조직이 서로 섞여있을 때 사람의 조직을 탐색하는 방법으로 유전자 증폭을 할 경우, digital PCR의 유효성에 대해 증명하였다.

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