• Journal of Veterinary Science
• /
• 제24권3호
• /
• pp.38.1-38.12
• /
• 2023

Background: Poor disease management and irregular vector control could predispose sheltered animals to disease such as feline Bartonella infection, a vector-borne zoonotic disease primarily caused by Bartonella henselae. Objectives: This study investigated the status of Bartonella infection in cats from eight (n = 8) shelters by molecular and serological approaches, profiling the CD4:CD8 ratio and the risk factors associated with Bartonella infection in shelter cats. Methods: Bartonella deoxyribonucleic acid (DNA) was detected through polymerase chain reaction (PCR) targeting 16S-23S rRNA internal transcribed spacer gene, followed by DNA sequencing. Bartonella IgM and IgG antibody titre, CD4 and CD8 profiles were detected using indirect immunofluorescence assay and flow cytometric analysis, respectively. Results: B. henselae was detected through PCR and sequencing in 1.0% (1/101) oral swab and 2.0% (1/50) cat fleas, while another 3/50 cat fleas carried B. clarridgeiae. Only 18/101 cats were seronegative against B. henselae, whereas 30.7% (31/101) cats were positive for both IgM and IgG, 8% (18/101) cats had IgM, and 33.7% (34/101) cats had IgG antibody only. None of the eight shelters sampled had Bartonella antibody-free cats. Although abnormal CD4:CD8 ratio was observed in 48/83 seropositive cats, flea infestation was the only significant risk factor observed in this study. Conclusions: The present study provides the first comparison on the Bartonella spp. antigen, antibody status and CD4:CD8 ratio among shelter cats. The high B. henselae seropositivity among shelter cats presumably due to significant flea infestation triggers an alarm of whether the infection could go undetectable and its potential transmission to humans.

• BMB Reports
• /
• 제38권6호
• /
• pp.683-689
• /
• 2005

Antibody to hepatitis B surface antigen (HBsAb) is the important serological marker of the hepatitis B virus (HBV) infection. Conventionally, the hepatitis B surface antigen (HBsAg) obtained from the plasma of HBV carriers is used as the diagnostic antigen for detection of HBsAb. This blood-origin antigen has some disadvantages involved in high cost, over-elaborate preparation, risk of infection, et al. In an attempt to explore the suitable recombinant HBsAg for the diagnostic purpose, the HBV S gene was expressed in Pichia pastoris and the product was applied for detection of HBsAb. Hepatitis B virus S gene was inserted into the yeast vector and the expressed product was analyzed by sodium dodecyl sulphate polyacrolamide gel electrophoresis (SDS-PAGE), immunoblot, electronic microscope and enzyme linked immunosorbent assay (ELISA). The preparations of synthesized S protein were applied to detect HBsAb by sandwich ELISA. The S gene encoding the 226 amino acid of HBsAg carrying ahexa-histidine tag at C terminus was successfully expressed in Pichia pastoris. The His-Tagged S protein in this strain was expressed at a level of about 14.5% of total cell protein. Immunoblot showed the recombinant HBsAg recognized by monoclonal HBsAb and there was no cross reaction between all proteins from the host and normal sera. HBsAb detection indicated that the sensitivity reached 10 mIu (micro international unit)/ml and the specificity was 100% with HBsAb standard of National Center for Clinical Laboratories. A total of 293 random sera were assayed using recombinant S protein and a commercial HBsAb ELISA kit (produced by blood-origin HBsAg), 35 HBsAb positive sera and 258 HBsAb negative sera were examined. The same results were obtained with two different reagents and there was no significant difference in the value of S/CO between the two reagents. The recombinant HBV S protein with good immunoreactivity and specificity was successfully expressed in Pichia pastoris. The reagent for HBsAb detection prepared by Pichia pastoris-derived S protein showed high sensitivity and specificity for detection of HBsAb standard. And a good correlation was obtained between the reagent produced by recombinant S protein and commercial kit produced by blood-origin HBsAg in random samples.

• Parasites, Hosts and Diseases
• /
• 제27권2호
• /
• pp.109-118
• /
• 1989

폐흡충 감염시 숙주 말초혈액에는 많은 항체가 생성되므로 각종 면역학적 진단에 이용되 고 있다. 그러나 이들 항체가 폐흡충 충체 구성 요소 중 주로 어느 기관(또는 구조물)에 대한 항체 인지에 대해서는 별로 연구된 바 없었다. 이 연구는 면역 조직화학적 방법을 이용하여 폐흡충 충체 의 부위별 항원성을 파악하기 위한 것으로 충체 절편 표본에서 흡반, 표피, 피극, 난황선, 장, 자성 및 웅성 생식기, 충란 등의 항원성을 비교 관찰한 것이다. 충체 표본으로는 감염 11∼20주 된 고양이의 폐조직에서 충낭(WOrm capsule)을 적출하여 포르말 린 고정 및 파라핀 포매한 것을 4㎛ 두께로 영아 사용하였고, 항혈청(1차 항체)으로 감염 11∼20 주 된 고양이 혈청을, 2차 항체로 peroxidase-conjugated goat anti-cat IgG를 사용하는 간접 면역 대소 염색법(indirect immunoperoxidase staining technique)을 이용하였고, 진한 황색 또는 황갈색으로 염색되면 양성으로 판정하였다. 항체의 희석 농도는 1차 항체 1 : 500~1 : 2,000, 2차 항체 1 : 200~1 : 500으로 하였고 10회 이상 반복 염색하였다. 실험 결과 장 상피의 표면(intestinal epithelial border), 장 내용물, 난황선(vitelline glands) 및 충낭 내의 충란(eggs in worm capsule) 등이 강한 양성 반응을 보였고 자궁 내 충란 및 충체 실질 조직 중 일부도 약하지만 양성 반응을 보였다. 한편 흡반(suckers), 표피(tegument), 피극(spines), 표피하세포(subtegumental cells), 장 상피세포의 세포질, 웅성 생식기관 및 난소 등은 음성 반응 을 나타내었다. 항원성의 강도를 순서대로 나열해보면 장 상피의 표면, 장 내용물, 충낭 내의 충란, 난황선, 자궁 내 충란, 충체 실질조직의 순이었다. 항원성이 강한 장 상피층 및 장 내용물은 1차 항체 1 : 4,000(2차 항체 1 : 200)에서도 양성 반응을 보였으나 충체 실질조직 중 일부는 1차 항체 1 : 500의 고농도(2차 항체 1 : 200)에서만 양성 반응을 보였다. 이상의 결과를 종합해 볼 때, 폐흡충 감염시 나타나는 혈청의 항체 반응은 충체의 배설물과 충 낭 주위에 산출된 충란에 의해 가장 강력히 유발되는 것이 아닌가 생각되며 이들이 충낭을 벗어나 숙주 조직으로 총수되는 가장 중요한 항원성 물질이 아닌가 추측된다.

• 한국식품영양과학회지
• /
• 제33권1호
• /
• pp.123-131
• /
• 2004

단백질의 Maillard 반응의 최종산물의 하나인 $N^{\varepsilon}$-car-boxy methyl lysine(CML)의 형성에 미치는 reactive oxygen species(ROS)의 영향을 살펴보았다. Glucose와 fructose는 자동산화과정을 통하여 CML 형성의 주된 propagator인 $\alpha$-dicarbonyl 화합물은 물론 glyoxal을 생성시키는 것이 확인되었다. 또한 이 과정은 ROS를 형성하는 천이금속이온에 의해 촉진되는 산화과정임을 알 수 있었다. 반응성이 높은 fructose에 비해 glucose의 경우가 ROS의 영향이 더욱 현저하게 나타났다. 불포화 지방산도 glyoxal을 형성하고 있었으며, 불포화도가 클수록 glyoxal 생성량이 빠르게 나타났으나 ROS의 영향은 비교적 작게 나타났다. Ascorbic acid 역시 ROS와는 무관하게 glyoxal을 생성하였다. 이는 ROS의 영향이전에 이들 물질들의 반응성이 매우 높다는 것을 의미하고 있다. Hippury lysine을 이용한 model system에서도 glucose로부터의 $N^{\varepsilon}$-carboxymethyl hipuryllysine(CMHL) 형성에서는 ROS의 영향이 높게 나타났으나, 반응성이 매우 높은 glyoxal에 의한 CMHL 생성에는 ROS의 영향이 거의 나타나지 않았다. CML에 특이적으로 결합하는 monoclonal antibody(6D12)를 이용한 antigen coated noncompetitive indirect ELISA 분석을 통해서 CML생성에 미치는 ROS의 영향을 살펴본 결과 대체로 위의 결과와 비슷한 경향을 나타내었다. 따라서 반응성이 높은 물질일수록 CML 형성에 ROS의 영향이 작게 나타남을 알 수 있었다.

• Parasites, Hosts and Diseases
• /
• 제26권2호
• /
• pp.73-86
• /
• 1988

간흡충(Clonorchis sinensis)의 조항원(WWA), 정제항원(ABA)의 민감도와 특이도를 비교해 보는 한편, 발육단계별로 충란 항원(EGA)-피낭유충 항원(MEA)-성충 항원(WWA)의 단백질 구성물질의 차이를 SDS-polyacrylamide gel electrophoresis(SDS-PAGE)로 관찰하고 이들 항원의 유용성을 효소면역반응(enzyme-linked immunosorbent assay: ELISA)으로 비교하여 다음과 같은 결과를 얻었다. 간흡충의 조항원으로부터 정제된 항원(ABA)은 항체와 결합한 결합항원(affinity-purified antibody(IgG) binding antigen:ABA)으로서 Ouchterlony 반응으로 간흡충 감염 가토혈청과 4개의 침모대 (WWA는 7개)를 형성하였다. WWA의 major protein들은 분자량 16,300∼18,500 및 28,000∼29,000 dalton의 Polypeptide band였고, ABA는 분자량 18,000∼21,000 및 29,000∼31,000 dalton의 단백질로 다소의 차이를 보였다. ELISA시첩에서 ABA는 WWA에 비해 현저히 민감도가 낮았다. WWA와 ABA에 대한 폐흡충 감염 사람혈청의 ELISA 정사에서 WWA는 Ouchterlony 양성자 8예중 3예에서 교차반응을 나타내었으나 ABA는 교차반응을 나타내지 않았다. 간흡충의 발육단계별 항원 단백질의 분자량 범위는 WWA 11,000∼80,000, MEA 15,000∼100,000, EGA 15,000∼200,000 dalton이었다. 주류원 단백질의 분자량은 EGA는 각각 36,600 (band No. 22), 38,500(No. 20), 64,000(No. 9), 62,000(No. 10), 54,500(No. 11), 53,000(No. 12) dalton, MEA는 각각 65,600(No. 3), 44,700 (No.7), 43,900(No. 8) dalton, WWA는 각각 16,300∼18,500(No. 31-32), 28,000∼29,000(No. 25), 11,000w13,000 (No. 35) 및 31,000(No. 24) dalton이었다. 즉, MEA와 EGA가 WWA보다 고분자의 단백질로 구성되어 있었다. 각 발육단계별 항원의 항원성은 ELISA 반응으로 볼 때 WWA가 가장 높았고, MEA는 약한 항원성만을 나타내었으며 EGA는 음성이었다. WWA와 간흡충 감염 가토혈청은 감염 4∼6주에 OD>1.0, 12주 이후 항체가의 plateau를 나타내었으나 MEA는 Ouchterlony 반응에서 EGA와 함께 음성반응을 보였다. ELISA에서도 중감염군(12∼20주)에서만 OD>0.6으로 미약한 항원성이 인정되었으며 경감염군은 전감염기간중 OD<0.6를 나타내었다. 이상의 결과로 보아 성충 항원이 가장 강력한 함원성 물질을 포함하고 있음이 명백하다고 생각된다. 그러나 성충 조항원은 면역진단에 사용할 겹우 타흡충과의 빈번한 교차반응이 예상되므로 민감도 및 특이도가 높은 정제 항원물질을 분리하는 일이 중요하다고 생각된다.

• 한국고분자학회:학술대회논문집
• /
• 한국고분자학회 2006년도 IUPAC International Symposium on Advanced Polymers for Emerging Technologies
• /
• pp.172-172
• /
• 2006

Molecular imprinting constitutes a valuable method of preparing polymeric materials with specific binding properties. The most conspicuous merit of molecular imprinting is that structurally three-dimensional recognition sites can be introduced into a polymer matrix with ease and low cost when compared with the complicated process of biological system for antigen and antibody. We used a thermally reversible bond for the preparation of the monomer-template complex, which allowed us to remove the template easily by means of a simple thermal reaction and to simultaneously introduce various functional groups into the cavity. This method is especially propitious for developing artificial receptors for molecules lacking strongly interactive groups.

• 생명과학회지
• /
• 제5권2호
• /
• pp.63-69
• /
• 1995

Cockroach antigen have been known as a cause of allergic disease. German ockroach(Blattella germanica L.) was chosen because it has the highest distribution range and poulation density. To identify the common and specific antigens of adult and larval stage of german cockroach, we made monoclonal antibodies which were confirmen by SDS-PAGE and EITB. Anti-B. germanica antibody producing hybridomas were 24 among the total 960wells. Only 4 hybridomas did not have cross reaction to other species of cockroach and hluse dust mites(Dermatophagodies farinae and D. pteronyssius). SDS-PAGE revealed about 20 bands from 90Kd to 15Kd to 15Kd. ETB showed specific antigens a6 60, 72 and 82Kd which were experimented by the culture supernatant of 4 selected hybridomas. Especially 60Kd coincided with a band of immunized mouse sera.

• 생명과학회지
• /
• 제5권2호
• /
• pp.7-7
• /
• 1995

Cockroach antigen have been known as a cause of allergic disease. German ockroach(Blattella germanica L.) was chosen because it has the highest distribution range and poulation density. To identify the common and specific antigens of adult and larval stage of german cockroach, we made monoclonal antibodies which were confirmen by SDS-PAGE and EITB. Anti-B. germanica antibody producing hybridomas were 24 among the total 960wells. Only 4 hybridomas did not have cross reaction to other species of cockroach and hluse dust mites(Dermatophagodies farinae and D. pteronyssius). SDS-PAGE revealed about 20 bands from 90Kd to 15Kd to 15Kd. ETB showed specific antigens a6 60, 72 and 82Kd which were experimented by the culture supernatant of 4 selected hybridomas. Especially 60Kd coincided with a band of immunized mouse sera.

• 센서학회지
• /
• 제25권5호
• /
• pp.320-325
• /
• 2016

In this paper, we propose an AlGaN/GaN-based extended-gate metal-insulator-semiconductor high electron mobility transistor (MISHEMT)-type biosensor for detecting streptavidin-biotin complexes. We measure the drain current of the fabricated sensor, which varies depending on the antibody-antigen reaction of streptavidin with biotin molecules. To confirm the immobilization of biotin polyethylene glycol (PEG) thiol, we analyze the Au surface of a GaN sample using X-ray photoelectron spectroscopy (XPS). The proposed biosensor shows higher sensitivity than Si-based extended-gate metal oxide semiconductor field effect transistor (MOSFET)-type biosensor. In addition, the proposed AlGaN/GaN-based extended-gate MISHEMT-type biosensor exhibits better long-term stability, compared to the conventional AlGaN/GaN-based MISHEMT-type biosensor.

• Journal of Multimedia Information System
• /
• 제9권2호
• /
• pp.161-170
• /
• 2022

Under General anesthesia with isoflurane, we insert a chicken's esophageal catheter into the near the left atrium. 1MHz radio wave was added to electrocardiogram electrodes of the esophagus, and the change of impedance (phase) was obtained by amplitude synchronous detection technique. At the same time, a thin tube is surgically inserted into the axillary artery to continuously measure blood pressure. The correlation between impedance (phase) and blood pressure was obtained. Both showed a very high correlation (R²=0.9665). It was also observed the waveform flowing from the left atrium into the left ventricle. When an individual infected with the avian influenza virus develops, the cytokine storms lead to hypotension earlier than the test for antigen-antibody reaction. In order to detect this, in the future, this impedance technique will be useful for screening individuals infected with avian influenza virus by measuring the blood pressure of chickens in cages in a non-contact manner using microwaves.