• 한국동물위생학회지
• /
• 제39권2호
• /
• pp.101-105
• /
• 2016

The present study investigated serological and molecular prevalence of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) infection in unvaccinated broiler breeder farms in Jeonbuk providence. An enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) had been used to determine antibody titers against MG and MS, and genome of these pathogens, respectively. Seventy five percent of farms were seropositive for MG and 94% of farms were seropositive for MS. In addition, the rate of antibody positive flocks against MG were 65.3% (32/49), while the rate of positive flocks against MS were 84.2% (80/95). The geometric mean antibody titers were $802.2{\pm}626$ and $27,726.7{\pm}2426$ against MG and MS, respectively. Interestingly, none of samples was positive for MG genome by PCR, while 94% (farms), 82% (flocks) and 62.6% (broiler breeder) were positive for MS genome by PCR. These findings suggest that the prevalence of MG or MS infection could be higher than expected. Thus, strict prevention program including vaccination and environmental sanitation should be implemented to avoid disease transmission from breeder to broilers as well as transmission among broilers.

• Bulletin of the Korean Chemical Society
• /
• 제33권5호
• /
• pp.1485-1490
• /
• 2012

An electrochemical immunoassay based on osmium-hippuric acid (HA) conjugate films onto the electrode is presented for the detection of urinary HA. This is the first report on the use of the oxidative electropolymerization of 5-amino-1,10-phenanthroline (5-$NH_2$-phen) for immobilizing an antigen, osmium-conjugated HA. As a redox mediator, [Os(5-amino-1,10-phenanthroline)$_2$(4-aminomethylpyridine-HA)Cl]$^{+/2+}$ (Os-phen-HA) was successfully synthesized and electropolymerized onto the screen-printed carbon electrodes (SPCEs). The interaction between osmium-HA conjugate films and antibody-HA ($anti$-HA) was performed by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The electrical signals were linearly proportional to urinary HA in the range of 0.1-5.0 mg/mL, which is sufficient for use as an immunosensor using a cutoff concentration of 2.0 mg/mL in urine samples. The proposed electrochemical immunoassay method can be extended to various applications for detecting a wide range of different small antigens in the health care area.

• Parasites, Hosts and Diseases
• /
• 제31권3호
• /
• pp.193-200
• /
• 1993

국내의 CUPtOSPOTidiUm 인체감염 실태를 조사하기 위하여 연세대학 세브란스 병원을 찾은 230명의 외래 환자 분변을 수거하였다. Acid-fast 염색, auramine-rhodamine 염색과 CWptosporidiwpowumoocyst에 특이적인 단클론 항체를 이용한 동정법을 이용하였다. 230명의 환자 중 48명(21%)이 AF 염색법에 의하여 50명(22%)이 AR 염색법에 의하여, 그리고 23명(10%)이 단클론 항체를 이용하는 형광현미경법으로 각각 Cryptosporidium에 감염된 것으로 조사되어 국내에서도 Cryptosporidium 인체 감염이 존재하고 있는 것으로 나타났다.

• 한국동물위생학회지
• /
• 제37권3호
• /
• pp.151-156
• /
• 2014

Porcine reproductive and respiratory syndrome virus (PRRSV) causes major economic losses to the Korean pig industry. ELISA tests using recombinant nucleocapsid protein of PRRSV have been most commonly used for PRRS diagnostics. In the current study, two commercial PRRSV ELISA kits (Bionote PRRSV Antibody ELISA and IDEXX 3XR PRRS Antibody ELISA) have been compared using sera collected from 19 swine farms in various stages of PRRSV infection confirmed by professional diagnostic centers. Thus 130 sera collected from 5 different farms with active PRRSV infection, 130 sera from 6 different farms with PRRS-stabilized status, and 140 sera from 8 different farms with PRRS-free status were evaluated to determine the correlation of test results between those ELISA kits. Both ELISA kits showed a good correlation [PRRSV-positive farms ($R^2$=0.6375) and stabilized farms ($R^2$=0.8928)] in sample-to-positive (S/P) ratio va lues. Among the 140 sera from negative farms, one sample was falsely positive by either of the ELISA kits. In conclusion, both of the ELISA kits showed a good correlation when applied on field samples collected from farms at various stages of PRRSV infection. Bionote ELISA or IDEXX ELISA gave a false positive result on 1 out of 140 negative samples so their specificity was calculated as 99.3%. Therefore, Bionote ELISA would be a good complementary and alternative method for IDEXX ELISA kit, and vice versa.

• Korean Chemical Engineering Research
• /
• 제49권4호
• /
• pp.387-392
• /
• 2011

생촉매인 효소의 기질선택성은 다양한 분야에서 유용하게 이용되고 있다. 그 중에서도 효소결합면역흡착검사(enzyme-linked immunosorbent assay, ELISA)는 항원항체의 결합을 항체와 공유결합된 효소의 반응으로 나타냄으로써 다양한 항원들의 진단을 가능케 했다. 하지만 기존의 효소결합면역흡착검사는 하나의 항체당 하나의 효소가 결합된 형태이기 때문에 감도(sensitivity)의 증가 폭에 그 한계가 있으며, 이를 극복하기 위한 방안으로 하나의 항체당 결합된 효소의 수를 증가시킴으로써 혁신적인 감도의 향상을 가져오는 연구가 진행되었다. 최근 나노바이오촉매(nanobiocatalyst, NBC) 접근방식을 이용한 효소활성의 안정화는 효소결합면역흡착검사의 감도 향상뿐만 아니라 그 성능의 안정성을 확보할 수 있는 연구결과로 이어지고 있다. 본 총설에서는 일반적인 효소결합면역흡착검사의 기본적인 원리와 감도향상을 위한 연구, 그리고 성능안정성(performance stability)을 향상시키기 위한 나노바이오촉매-결합면역흡착검사(Nanobiocatalyst-Linked Immunosorbent Assay, NBC-LISA)에 대하여 살펴보고자 한다.

• 한국어병학회지
• /
• 제24권2호
• /
• pp.47-51
• /
• 2011

The present study demonstrated lymphocystis disease virus (LCDV) propagation through cytopathic effects (CPE) formation and LCDV detection in olive flounder fin (FFN) cells by polymerase chain reaction (PCR) and fluorescent antibody technique (FAT) methods. Tissue filtrates from the cluster cells produced CPE in FFN cells, which initially cells became enlarged and gradually underwent fusion en masse. Infectivity of culture grown LCDV using the FFN cells reached $10^{2.3}$ $TCID_{50}$/ml at 4 days post infection and the highest titer was measured $10^{6.5}$ $TCID_{50}$/ml at 12 days. The viral DNA was detected in the cell culture supernatants showing CPE and the CPE cells by PCR. Antigen specific strong fluorescence reacting with monoclonal antibody against the virus revealed the presence of viral antigen in the cytoplasm of infected FFN cells. These results suggest that the FFN cell line originated from the olive flounder has a susceptibility of the LCDV.

• BMB Reports
• /
• 제29권3호
• /
• pp.192-199
• /
• 1996

A rapid and sensitive method has been developed to detect a-fetoprotein (AFP) in human serum by a two-site sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) for human AFP within 1 h. To obtain the most sensitive and reliable MAbs. 12 kinds of MAbs (HPJ1 to HPJ12) as a capture antibody and 4 kinds of horseradish peroxidase (HRP) conjugated MAbs as a tracer antibody were investigated. Among these, only HPJ 10-HRP conjugated HPJ 1 (HPJ 10-HPJ $1^*$) and HPJ 11-HRP conjugated HPJ 10 (HPJ 11-HPJ $10^*$) were chosen as candidates based on the linearity of the standard curve and the sensitivity of the assay. To further characterize these two pairs. MAbs against human AFP were purified from hybridoma cells. conjugated with HRP. and then characterized to optimize the two-site sandwich ELISA The HPJ 10-HPJ $1^*$ pair showed a sensitivity of 1 ng/ml and a better reproducibility than the HPJ 11-HPJ $10^*$ pair when the human sera were incubated at $37^{\circ}C$ for 30 min. The results obtained for 480 randomly selected human sera showed 0~20 ng/ml of AFP values for the normal human sera. To test the utility of our kit, AFP concentrations were determined for 951 human sera (including 85 normal sera, 480 random blood sera, 213 HBsAg-positives. 50 anti-HCV antibody positives. and 47 malignant diseases) and compared with other commercially available AFP detecting kits. These results show that the present two-site sandwich ELISA method is a rapid, sensitive, and reliable procedure for detecting AFP in human serum.

• 대한수의학회지
• /
• 제32권1호
• /
• pp.57-61
• /
• 1992

Borna disease(BD) virus 특이항체검출에 대한 세 가지 혈청진단법(간접형광항체법, 세포효소면역반응법, 혈청중화시험)의 정확도를 비교하기 위해 BD virus를 실험적으로 감염시킨 273수의 토끼의 혈청으로 시험하였다. 혈청중 간접형광항체법에 의하여 판정된 123혈청들은 모두 세포효소면역반응법에 의해서 양성으로 판정되었으나 혈청중화시험법에 의해서는 단지 27혈청에서만 양성으로 판정되었다. 혈청중화시험법은 간접형광항체법과 세포효소면역반응법에 비하여 민감도가 훨씬 낮게 나타나 BD virus의 혈청학적 연구에는 간접형광항체법 및 세포효소면역반응법의 활용도가 높을 것으로 생각된다.

• 한국동물위생학회지
• /
• 제37권3호
• /
• pp.143-150
• /
• 2014

Porcine reproductive and respiratory syndrome virus (PRRSV) is the etiological agent of PRRS characterized by reproductive losses in sows and respiratory disorders in piglets. The PRRSV is a small enveloped virus containing a positive-sense, single-stranded RNA genome and divided into two genotype, type 1 (European) and type 2 (North American), respectively, by nucleotide identity. In this study, ORF7 gene of the type 1 and type 2 PRRSV was cloned and expressed in Baculovirus expression system. Also, monoclonal antibodies (MAbs) against ORF7 were produced and characterized. The expressed ORF7 proteins in the recombinant virus were confirmed by indirect fluorescence antibody (IFA) test using His6 and PRRSV-specific antiserum. A total of eight MAbs were produced and characterized. One (3G12) MAb was type 1 PRRSV ORF7-specific and two (6B10 and 16H8) were type 2 PRRSV ORF7-specific. Other five (1A1, 2A4, 4B4, 12C4 and 13F11) MAbs reacted with both type 1 and type 2 PRRSV. Some PRRSV ORF7-specific MAbs recognized the porcine tissues infected with PRRSV by IFA or immunohistochemistry (IHC) assay. From this experiment, it was confirmed that MAbs produced in this study were PRRSV ORF7-specific and could be used as reliable reagents for type 1/type 2 PRRSV detection.

• 한국동물위생학회지
• /
• 제33권2호
• /
• pp.105-111
• /
• 2010

Rabies virus which belongs to the genus Lyssavirus of the family Rhabdoviridae is known as a highly neurotropic virus and causes fatal encephalitis accompanied by severe neurological symptoms in almost all mammals, including humans. In this study, monoclonal antibodies (MAbs) against rabies virus were produced, characterized and applications of MAbs as diagnostic reagents were assessed Spleen and inguinal lymph node cells from Balb/c mouse immunized with purified rabies virus were fused with SP2/O myeloma cells using polyethylene glycol (PEG) and hybridoma cells producing rabies virus-specific MAbs were screened by an indirect fluorescent antibody test. A total of ten MAbs were produced against rabies virus. The protein specificity and neutralizing activity of MAbs were determined by Western blot analysis and fluorescent antibody virus neutralization test, respectively. As a result, two MAbs, 5G3 and 6H4 had specificity for nucleoprotein (N protein) and two other MAbs, 5B1 and 5C1 had neutralizing activity for rabies virus. Some MAbs recognized the rabies virus-infected bovine brain stem cells by immunohistochemistry (IHC) assay. In conclusion, it was confirmed that MAbs produced in this study were rabies virusspecific and could be used as reliable diagnostic reagents for the detection of rabies virus.