• 대한바이러스학회지
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• 제28권1호
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• pp.31-38
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• 1998

In Korea, all domestic made test systems for detecting antibodies in HIV-1 contain the antigens from human immunodeficiency type 1 (HIV-1) subtype B. However, because HIV-1 subtype O is significantly different in amino acid sequences from all other subtypes of HIV-1, there has been a need for developing a test for detecting antibodies in subtype O. For this purpose, the entire nucleotide sequence corresponding to the extracellular domain of the transmembrane glycoprotein of HIV-1 subtype O was synthesized with consideration of Escherichia coli condon usage. Various regions of the extracellular domain were cloned into E. coli expression vectors and tested for levels of protein production. The nucleotide sequence, named ECTM, that can encode a 129 amino acid-long peptide, was found to be expressed at a high level in E. coli. The protein of approximately 17 kDa specifically reacted with sera from individuals infected with HIV-1 subtype O. The ECTM protein was purified to near homogeneity by the CM-T gel chromatography, using concentrated, denatured inclusion bodies. In Western blot analysis, the purified viral antigen reacted with sera from individuals infected with subtype O more efficiently than subtype B. The enzyme linked immunoabsorbent assay (ELISA) system was developed using the subtype O viral protein and compared with the commercially available kit lacking the antigens from subtype O. The ELISA kit containing the subtype O antigen ECTM alone efficiently reacted with sera from individuals infected with subtype O. The subtype O antigen-containing kit produced a positive absorbence even when sera were diluted 512-fold, suggesting a high sensitivity. The commercially available kit also reacted with subtype O sera, but produced a negative result at a dilution of 8-fold. Our results suggest that the currently available kit may not be able to efficiently detect subtype O sera and that the viral protein developed in this study may be added to the current system to maximize the detection of sera from individuals infected with subtype O.

• 대한의생명과학회지
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• 제3권2호
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• pp.107-114
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• 1997

전립선특이 항원 (PSA: Prostate Specific Antigen)은 전립선 세포에서 분비되는 단백질로서 장기 특이성이 높아 전립선암의 조기진단 및 치료의 예후를 판단하기 위해 임상에서 널리 사용되고 있는 혈청 종양표지검사의 대상항원이다. 그러나 혈청 PSA치는 전립선암 뿐 아니라 양성 전립선질환인 전립선비대증 (BPH), 전립선 경색, 전립선 염증 등에서도 상승될 수 있다. 혈청 내의 PSA는 여러 가지 분자형태로 이루어져 있으며 대표적으로 ${alpha}_1$-antichymotrypsin과 결합된 결합형 PSA(PSA-ACT)와 유리형 PSA (free PSA)로 나눌 수 있다. 전립선 암 환자의 혈청에는 결합형 PSA가 95% 이상으로 매우 높고 유리형 PSA의 분포는 매우 작은 반면 전립성비대증에서는 유리형 PSA의 농도가 높아지므로, 결합형 PSA와 동시에 유리형 PSA를 측정하면 종양표지검사로서의 특이성과 예민도를 높일 수 있다는 보고가 있어, 최근 혈청내의 유리형 PSA의 측정의 중요성이 임상적으로 대두되고 있다. 이에 본 연구에서는 sandwich 원리로 유리형 PSA 효소면역 측정법 (EIA) kit를 개발하고 그 유용성을 검토한 결과, 고, 저 농도에서의 일내, 일간 변이 계수 (CV)는 4% 이하였으며 상품화된 free PSA kit와 비교하였을 때 두 방법간의 상관계수는 0.9965으로 매우 양호하였다. 또한 농도가 높은 세 환자의 검체를 희석하여 직선성 검사를 하였을 때 상관계수가 모두 0.995이상으로 나타났다. 또한 microplate 법에서 문제될 수 있는 hook effect는 유리형 PSA농도가 40 ng/mL까지 나타나지 않았으며, 98.9%~104.1%의 회수율을 나타내었다. 따라서 본 연구에서 개발한 면역측정법 kit는 혈청 중의 유리형 PSA를 정확하고 간편하게 분석할 수 있으므로 임상 실험실에서 전립선암이나 전립선질환의 진단 및 치료효과의 판정에 유용하게 사용될 수 있을 것으로 기대된다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7433-7438
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• 2013

Aim: To identify new biomarkers for NPC diagnosis with an anti-EBV Western blot test kit. Methods: Serum samples from 64 NPC patients and healthy subjects with four specific VCA-IgA/EA-IgA profiles were tested with an anti-EBV Western blot test kit from EUROIMMUN AG. Proteins were quantified with scores of intensity visually assigned to the protein bands. The markers which showed statistical differences between the NPC and non-NPC subjects were further evaluated in another 32 NPC patients and 32 controls in comparison with established biomarkers including VCA-IgA, EA-IgA, EBV-related protein IgG, and EBV DNA. Results: Among the markers screened, EA-D p45-IgG showed a statistically significant difference (p < 0.05) between NPC and non-NPC subjects with VCA-IgA positivy. In 32 VCA-IgA positive NPC patients and 32 control subjects, the diagnostic accuracy of EA-D p45-IgG was 78.1% with a positive predictive value of 77.8% and a negative predictive value of 78.6%. In the verification experiment, the specificity and sensitivity of EA-D p45-IgG were 75.0% and 90.6 %, respectively. Conclusions: EA-D p45-IgG might be a potential biomarker for NPC diagnosis, especially among VCA-IgA positive subjects.

• 대한수의학회지
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• 제42권4호
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• pp.545-550
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• 2002

The raccoon dog (Nyctereutes procyonoides) may be infected by Dirofilaria immitis. However, there has been no report on dirofilarial infection in the raccoon dog in Korea. In this study, we report on D. immitis infection in two wild raccoon dogs captured in the Daejeon area. The two raccoon dogs were referred to the Veterinary Teaching Hospital, Chungnam National University for diagnosis of D. immitis infection. The modified Knott's test for the detection of blood D. immitis microfilariae was positive, and serological test (FASTest$^{(R)}$ HW Antigen ELISA kit, Diagnostik Mega Cor, Austria) for D. immitis was positive as well. Additionally, D. immitis microfilariae were differentiated from other microfilariae by using acid phrnphatase histochemical staining (Leucognost-SP$^{(R)}$kit, Diagncstica MERCK, Germany). The two raccoon dogs were necropsed and D. immitis infection was confirmed.

• Parasites, Hosts and Diseases
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• 제51권3호
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• pp.375-377
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• 2013

In the present study, we first report the seroprevalence of Dirofilaria immitis in dogs in Shenyang, northeastern China. Sera from 528 randomly selected dogs were examined for D. immitis antigen using SNAP$^{(R)}$4Dx test kit; 12.7% tested showed seropositive. No significant difference of infection was observed in different genders and breeds (P>0.05), but the difference was significant in different age groups and rearing conditions (P<0.05). The result suggested that the risk of exposure to D. immitis in dogs is high in Shenyang, and should be given attention.

• 대한수의학회지
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• 제38권3호
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• pp.595-599
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• 1998

The present study was undertaken to develop the kit for indirect immunofluorescence antibody test(IFAT) and to investigate the prevalence of N caninum in cattle and dogs in Korea. The neo-antigen kit of IFAT developed in our laboratory was proved diagnostic efficacy by compared with the kit of veterinary medical research and development(VMRD). A survey of N caninum infections among cow and dogs in elevn areas of southern part of country was performed using a IFAT. The infection rate of 190 nationwide cattle was 8.4%(16/190) but was 75%(45/60) at P area in Chunnam province where the cattle showed the abortion repeatedly. Any of the dogs was not N caninum -positive in the Kangwon and Kyonggi areas.

• 한국수산과학회지
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• 제48권2호
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• pp.158-167
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• 2015

A lateral flow immunoassay kit based on antigen-antibody interactions was developed to detect residues of beta-lactams, quinolones, tetracyclines, and sulfonamides in farmed fish. Group-specific antibodies showing cross-reactivity with other antibiotics in the same group were produced in rabbits. The rabbits were immunized eight times to obtain the maximum titers. Antibodies were extracted from the antisera collected from the immunized rabbits and produced group-specific reactions with antibiotics from the four groups. A kit was prepared that optimize conditions for the antigen-antibody reaction, using colloidal gold conjugated antibodies, and was designed to detect the four groups of antibiotics simultaneously. The kit enabled the detection of antibiotics in the four groups at below maximum residue limits (MRLs), which were $200{\mu}g/kg$ for tetracyclines, $100{\mu}g/kg$ for sulfonamides, $50{\mu}g/kg$ for beta-lactams, and $100{\mu}g/kg$ for quinolones. The cross-reactivity of the antibodies ranged from 10-80% for the sulfonamides, 20-100% for tetracyclines, 38-100% for quinolones, and 20-100% for the beta-lactams, confirming that the antibodies were group specific. The test kit was used 30 times to examine spiked antibiotics at the limits of detection (LODs) and all produced positive results, indicating high sensitivity. The LODs for the assay ranged from 4-20 ng/mL for beta-lactams, 25-50 ng/mL for sulfonamides, 20-100 ng/mL for tetracyclines, and 30-80 ng/mL for quinolones, and there were no false negative reactions at above these LODs. In addition, all of the LODs of the developed kit were correlated with high-performance liquid chromatography (HPLC) data. Our lateral flow immunoassay kit can simultaneously detect antibiotic residues from a large number of fish samples rapidly, strengthening the safety of domestic farmed and imported fish.

• Tuberculosis and Respiratory Diseases
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• 제53권4호
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• pp.389-400
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• 2002

배 경 :결핵의 조기치료와 전염방지를 위해서 신속하고 간편한 결핵진단법의 개발이 요구되고 있는 실정에서 결핵균에 특이한 38-kDa단백을 포함하여 금 콘쥬게이트에 결합된 유전자재조합 항원을 혈청과 반응시켜 항결핵 항체를 발견하도록 고안된 카드형태의 혈청학적 진단기법인 Xeniss Rapid TB kit가 결핵의 진단에 유용하게 이용될 수 있는 지를 알아보고자 하였다.방 법 :188명의 결핵환자(폐결핵 177명, 폐외결핵 11명)와 82명의 접촉자, 그리고 57명의 건강한 성인을 대상으로 하였으며, 연구대상자의 혈청을 이용하여 Xeniss Rapid TB kit의 민감도, 특이도, 양성예측율, 그리고 음성예측율을 조사하였다. 결 과 : 전체적인 민감도는 73.9%, 특이도 81.3%, 양상예측율 84.2%, 그리고 음성예측율은 85.8%였다. 진단시점부터 검사시점간의 시간간격에 따라서는 1개월 이내에서 61.5%로 가장 낮고 점차 증가하여 4-6개월 시점에 94.4%로 가장 높았으며 이후 점차 감소하여 12개월 이상 경과한 시점에서는 67.4%의 양성반응율을 보였다. 페외결핵 환자(90.9%)에서는 폐결핵 환자(72.8%)보다 양성반응율이 높았다. 객담도말양성 (76.2 대 68.4%), 방사선 사진상 중증 (79.3 대 63.3%), 공동성 병소(75.7 대 70.0%), 과거 치료력 (76.3 대 73.3%)이 있는 환자군에서 상대적으로 높은 양성반응율을 보였으며, 당뇨병을 동반한 환자군(69.0 대 74.8%)과 노인환자군(68.l 대 100%)에서는 상대적으로 낮은 양성반응율을 보였다. 건강성인군 7.0%, 환자가족군 17.5%에 비해 병원직원군에서 57.9%의 양성반응율을 보여 활동성 결핵환자와 장기간 지속적으로 접촉한 군에서 높은 양성반응율을 보였다. 결 론 : Xeniss Rapid TB kit는 신속하고 간편하며, 민감도와 특이도가 비교적 높고 특히, 폐외결핵에서는 높은 양성반응율을 보여 폐결핵, 폐외결핵, 그리고 감염자의 진단에 보조적 검사법으로 유용하게 이용될 수 있을 것으로 생각된다.

• Journal of Oral Medicine and Pain
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• 제25권2호
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• pp.167-172
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• 2000

Both immune reaction and hypersensitivity reaction are occurred by the same mechanism, the antigen and antibody reaction. The favorable result of this reaction towards a host is called clinically an immune reaction and the opposite results is called an hypersensitivity reaction. Type IV hypersensitivity reaction is a delayed type which is related to the cellular immune reaction and a contact hypersensitivity is included in this type. Various dental materials such as metal (mercury, nickel, chrome, cobalt), resin and eugenol are etiologic substances. Patch test kit is composed of test substance with a controlled concentration which respond only to a susceptible patient and an aluminum chamber, and etiologic substances for hypersensitivity can be easily and comfortably found just by applying the kit to the patient's skin. In this case report, the patch test was performed to a patients with oral lichen planus and the allergen, restorative material was found. After removal of the matching restoration from the patient's mouth, the symptom was improved.

• 대한의생명과학회지
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• 제21권1호
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• pp.9-14
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• 2015

Mouse hepatitis virus (MHV) is a major pathogen in laboratory mice that usually leads to fatal diseases, such as hepatitis, multiple sclerosis, encephalitis, and respiratory disease. MHV has a high infection rate, and it needs to be detected as soon as possible to prevent its spread to other facilities. However, MHV detection by enzyme-linked immunosorbent assay (ELISA) often gives false positives; thus, it is very important that the results are confirmed as true positives in the early infection stage or distinguished as false positives with more accurate, reliable methods. Under microbiological screening, MHV ELISA-positive mice were found in four GFP-tagging transgenic mice. To verify the detection of the MHV antigen directly, reverse transcription polymerase chain reaction (RT-PCR) was performed, and the mice were determined to be MHV negative. Additional serum antibody-based screening was conducted with three different ELISA kits, and multiplexed fluorometric immunoassay (MFIA) was performed to confirm their accuracy/sensitivity. In brief, the ELISA kit for A59 nucleocapsid protein (MHV-A59N) revealed MHV ELISA positivity, while other ELISA kits (MHV-S lysate and MHV-JHM lysate) demonstrated MHV negativity. In MFIA, only the test for the recombinant A59 nucleocapsid antigen was MHV positive, which was consistent with the ELISA results. These results suggest that the ELISA kit with the recombinant A59 nucleocapsid antigen might induce non-specific MHV ELISA positivity and that confirmation is therefore essential.