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소의 임상병리 가검물에서 Mycobacterium species 감별진단을 위한 multiplex PCR 기법
김용환,조호성,강성귀,조경오,박형선,이봉주,박남용,Kim, Yong-hwan,Al-Haddawi, MH,Cho, Ho-seong,Kang, Sung-kwi,Cho, Kyoung-oh,Park, Hyung-seon,Lee, Bong-joo,Park, Nam-yong 대한수의학회 2001 大韓獸醫學會誌 Vol.41 No.4
A multiplex PCR technique was developed for detecting specifically each Mycobacterium bovis, M. tuberculosis, M. avium and M. avium subsp, paratuberculosis, respectively, using clinical samples of field cattle. To apply this novel technique to clinical specimens, blood sample was obtained from live cows comprising 11 intradermal tuberculin test (ITT)-positive and 17 ITT-negative and tested by multiplex PCR. Positive results were obtained from 15 cows by the multiplex PCR, showing that 4 (23.5%) of the 17 ITT-negative cows were multiplex PCR positive. The multiplex PCR results also showed that among the 15 positive cows, 7 (46.7%) were infected with M. bovis, 1 (6.7%) with M. tuberculosis and 7 (46.7%) with M. avium. The sensitivity and specificity of multiplex PCR in comparison with those of ITT were 100% and 76.5%. The correlation between the multiplex PCR and ITT assays with blood samples was considered excellent, 85.7% agreement and ${\kappa}=0.72$. The results obtained, using reference mycobacterial strains and typed clinical samples, show that the multiplex PCR method may be a rapid, sensitive, and specific tool for the differential identification of various mycobacterial strains in a single-step assay. Therefore, multiplex PCR assay is a useful tool for early diagnosis of tuberculosis in live cattle and to identify the species or complex of mycobacterium from clinical samples.
In situ RT-PCR 및 In situ hybridization 기법에 의한 닭 뉴캣슬병의 진단법 개발
박남용,최효임,조호성,강성귀,조경오,Park, Nam-Yong,Choi, Hyo-Im,Cho, Ho-Seong,Kang, Sung-Kwi,Cho, Kyoung-Oh,Brown, Corrie 대한수의학회 2002 大韓獸醫學會誌 Vol.42 No.3
Newcastle disease (ND) is a highly contagious infection of poultry, Two pathology-based techniques, in situ RT-PCR and in situ hybridization (ISH) were applied to formalin-fixed, paraffin-embedded tissues from chickens naturally infected with velogenic ND virus (VNDV). Two pairs of primers and a probe for ISH and in situ RT-PCR, respectively, were selected from highly conserved region of matrix gene of NDV. The ISH experiment was carried out using MicroProbe$^{TM}$ capillary action system within 2 hours. In situ RT-PCR was performed using MicroProbe$^{TM}$ capillary action system and GeneAmp In Situ PCR system. With ISH and in situ RT-PCR, viral nucleic acid was detected in the central nervous system of chickens from infected with neurotropic velogenic Newcastle disease virus (NVNDV), whereas viral nucleic acid was detected in various organs or tissues of chickens from infected with viscerotropic velogenic Newcastle disease virus (VVNDV). In the NVND group, positive signals were characteristically defined in the cytoplasm of neuron, vascular endothelial cells, and perivascular mononuclear macrophages in the central nervous system. One of NVND group, chicken from one farm exhibited positive signals in the bronchial epithelium. The VVND group chickens showed positive reaction in the macrophages, vascular endothelium, and bronchiolar epithelium. Markedly, viral nucleic acid was detected in the macrophages of morphologically normal tissues which were peripheral or located in distant areas from lesions. The central nervous system of chickens infected with VVND virus had positive signals in the vascular endothelial cell, perivascular mononuclear macrophages and some neuron. The number and intensity of the positive cells by in situ RT-PCR were more and stronger, respectively, in comparison with those by ISH. Particularly, positive reaction was detected in macrophages infiltrating in cardiac muscle by in situ RT-PCR, but not obtained by ISH. Therefore, these results demonstrated that ISH is a rapid diagnostic method for detection of NDV and in situ RT-PCR can be used as an efficient method for detection of low viral load infection or subclinical viral infection of NDV.
Physostigmine과 procyclidine으로 구성된 유기인제 중독 복합예방제에 대한 2주 반복투여 용량설정시험
황재식(Zai-Zhi Huang),강현구(Hyun-Gu Kang),박선희(Sun-Hee Park),강성귀(Seong-kwi Kang),이종성(Jong-Sung Lee),박종일(Jong-Il Park),김왕수(Wang-Soo Kim),피택산(Taek-San Phi),황석연(Seok-Yeon Hwang),강종구(Jong-Koo Kang),김윤배(Yun-Bae 한국실험동물학회 2003 Laboratory Animal Research Vol.19 No.4