Abstract:
The present invention relates to a detection method using magnetic nanoparticles for rapidly isolating Mycobacterium avium subsp. paratuberculosis from fecal samples and, more specifically, to a method for detecting the Mycobacterium avium subsp. paratuberculosis which isolates the Mycobacterium avium subsp. paratuberculosis with high sensitivity which can be existed in animal fecal samples using the magnetic nanoparticles to which Mycobacterium avium subsp. paratuberculosis specific antibodies are combined and rapidly and simply checks infection of the Mycobacterium avium subsp. paratuberculosis by performing polymerase chain reaction using primers which are specifically combined to DNA of the Mycobacterium avium subsp. paratuberculosis. According to the present invention, the detection method using the magnetic nanoparticles for rapidly isolating the Mycobacterium avium subsp. paratuberculosis from fecal samples can easily detect the Mycobacterium avium subsp. paratuberculosis even if small number of the Mycobacterium avium subsp. paratuberculosis is existed in biological samples such as feces and prevent contagion to different entities by isolating the infected entities from an allied army by performing the method simply and rapidly.
Abstract:
PURPOSE: A method for detecting a quinolone resistant Campylobacter variant in a biological sample and a primer set and a kit for PCR for the same are provided to cheaply and simply detect and distinguish quinolone resistant variants of C. jejuni and C.coli. CONSTITUTION: A method for distinguishing and detecting a wild type and a quinolone resistant variant type of C. jejuni and C.coli in a biological sample comprises: a step of isolating DNA from the biological sample; a step of performing PCR using the DNA as a template and a primer set for PCR containing an oligonucleotide of sequence numbers 1, 2, 3, 4, 5, and 6; and a step of analyzing a PCR product and distinguishing wild type and variant type.
Abstract:
PURPOSE: Diagnostic methods of foot-and-mouth disease using recombinant 3ABC non-structural protein expressed in insect cells and a monoclonal antibody is provided, thereby more rapidly and accurately diagnosing the foot-and-mouth disease than the prior methods. CONSTITUTION: A gene encoding a recombinant 3ABC non-structural protein derived from Korean foot-and-mouth disease virus O/SKR/2000 has the nucleotide sequence of SEQ ID NO: 1. A recombinant baculovirus to be expressed in insect cells is prepared by co-transfection with a recombinant vector containing the recombinant 3ABC non-structural protein gene of SEQ ID NO: 1. The recombinant 3ABC non-structural protein is expressed by infection of the insect cells with the recombinant baculovirus. A hybridoma cell line 3F-11(KCTC 10138BP) is prepared by introducing the recombinant 3ABC non-structural protein expressed in E. coli into an animal, collecting an immunized cell from the animal and fusing the immunized cell with a cancer cell. A monoclonal antibody is produced from the hybridoma cell line 3F-11(KCTC 10138BP). A diagnostic method of foot-and-mouth disease comprises the steps of: (1) diluting the recombinant 3ABC non-structural protein in a coating buffer solution and pouring the diluate on a plate; (2) reacting the testing serum with the plate; (3) reacting a conjugate, which binds with an antibody for foot-and-mouth disease virus in the testing serum and has an enzyme, a radioactive material or a fluorescent material, with the testing serum; and (4) measuring intensity of the enzyme reaction, fluorescence reaction or radiation reaction with the conjugate.
Abstract translation:目的:提供使用在昆虫细胞和单克隆抗体中表达的重组3ABC非结构蛋白的口蹄疫诊断方法,从而比现有方法更快速和准确地诊断口蹄疫。 构成:编码来源于韩国口蹄疫病毒O / SKR / 2000的重组3ABC非结构蛋白的基因具有SEQ ID NO:1的核苷酸序列。在昆虫细胞中表达的重组杆状病毒由 用含有SEQ ID NO:1的重组3ABC非结构蛋白基因的重组载体共转染。通过用重组杆状病毒感染昆虫细胞表达重组3ABC非结构蛋白。 通过将在大肠杆菌中表达的重组3ABC非结构蛋白导入动物中,从动物中收集免疫的细胞并将免疫的细胞与癌细胞融合来制备杂交瘤细胞系3F-11(KCTC 10138BP)。 从杂交瘤细胞系3F-11(KCTC 10138BP)产生单克隆抗体。 一种口蹄疫诊断方法,包括以下步骤:(1)将重组3ABC非结构蛋白稀释于包被缓冲液中,并将稀释液倒入平板中; (2)使测试血清与平板反应; (3)使与试验血清中的口蹄疫病毒抗体结合的缀合物与测试血清具有酶,放射性物质或荧光物质; 和(4)测量酶反应的强度,荧光反应或与缀合物的辐射反应。
Abstract:
본 발명은 개 부루셀라병의 진단에 있어서, 기존의 개발된 혈청학적 진단법의 단점인 부루셀라(Brucella) 균의 점조성(mucoid )으로 인한 자가응집반응 및 교차반응을 줄이기 위하여 부루셀라 케니스 변이균주( Brucella canis mutant strain Mucoidless)를 이용한 개 부루셀라병의 신속평판응집반응 진단킷트에 관한 것이다. 부루셀라 케니스 변이균주는 부루셀라 케니스 균주( Brucella canis strain)에 비하여 병원성 감소, 비특이적인 응집성 감소, 높은 소수성 등에서 큰 차이를 나타내고, PH 4∼6의 범위에서 빠른 시간에 응집하는 결과를 나타내어 현재 피트만 모르사에서 사용하고 있는 부루셀라 오비스 ( B. ovis ) 균주보다도 훨씬 신속하게 반응하는 결과를 나타내어 응집반응 원리를 이용한 혈청학적 진단시 비특이반응을 줄이는데 매우 효율적이다. 부루셀라 케니스 변이균주를 이용한 신속평판응집반응법은 진단효율도 높을 뿐 아니라 1시간이내에 검출할 수 있을 정도로 신속하고, 사용방법도 간편함을 특징으로 한다.