• Tuberculosis and Respiratory Diseases
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• 제49권5호
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• pp.558-567
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• 2000

배경 : 현행 도말 및 배양 등의 미생물학적 검사법의 제한점을 보완할 수 있는 신속하고 간편한 결핵진단 방법의 하나로서 가장 많이 연구되어 온 분야가 혈청학적인 방법이다. 이 중 현재까지 결핵의 혈청학적 진단에 유용성이 높은 것을 평가되는 항원의 하나가 38-kDa으로 대표되는 결핵균 분비항원이다. 이 38-kDa을 주항원 성분으로 하여, 간편하게 실험할 수 있도록 kit화된 수입제품이 국내에서 널리 시판되고 있는 실정에서 국내의 회사에서 개발한 ELISA kit(Erum Biotech Co.)를 이용하여 폐결핵의 혈청학적 진단이 얼마나 유용한 지를 평가하고자 하였다. 방법 : 도말 및 배양검사장 균양성으로 진단된 후 항결핵치료를 받고 있는 폐결핵환자 333명(검사당시 균양성 환자 212명, 균음전된 환자 121명), 건강 성인 80명, 그리고 국립마산결핵병원에서 1년 이상 근무하며 환자와 접촉을 자주 하게되는 접촉군 61명 등 총 474명을 대상으로 하여 결핵의 혈청학적 진단용 ELISA kit를 이용하여 시험하였다. 결과 : 1) 균양성 활동성 폐결핵환자 212명에 대한 ELISA kit의 양성반응률은 82.1%, 균음성 활동성 폐결핵환자 121명에 대한 양성반응률은 73.6%로 이 두 군 사이에는 통계학적으로 유의한 차이가 없었다(p>0.05). 2) 접촉 대조군 61명에 대한 양성반응률은 14.8%, 건강 대조군 80명에 대한 양성반응률은 2.5%로 이 두 군 사이에는 통계적으로 유의한 차이가 있었다(plt;0.001). 3) 활동성 폐결핵환자 333명 모두에 대한 양성반응률은 78.90%, 대조군 141명 모두에 대한 양성반응률은 7.8%로 이 두 군 사이에는 통계적으로 유의한 차이가 있었다(p<0.001). 4) ELISA kit의 민감도는 78.9%, 특이도는 97.5%였으며, 유병율이 60.1% 수준일 때의 양성예측율은 96.1%, 음성예측율은 65.0%였다. 결론 : ELISA kit는 민감도나 특이도 면에서 수입시판되고 있는 ICT와 비교할 때 비슷한 결과를 보이며, 전통적으로 결핵을 진단하는 데 사용하던 흉부 X-선 사진, 항산균 염색 및 배양 등과 함께 보조적인 도구로 사용 할 수 있을 것으로 생각된다.

• Parasites, Hosts and Diseases
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• 제54권3호
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• pp.375-380
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• 2016

Angiostrongyliasis is difficult to be diagnosed for the reason that no ideal method can be used. Serologic tests require specific equipment and are not always available in poverty-stricken zone and are time-consuming. A lateral flow immunoassay (LFIA) may be useful for angiostrongyliasis control. We established a LFIA for the diagnosis of angiostrongyliasis based on 2 monoclonal antibodies (mAbs) against antigens of Angiostrongylus cantonensis adults. The sensitivity and specificity were 91.1% and 100% in LFIA, while those of commercial ELISA kit was 97.8% and 86.3%, respectively. Youden index was 0.91 in LFIA and 0.84 in commercial ELISA kit. LFIA showed detection limit of 1 ng/ml of A. cantonensis ES antigens. This LFIA was simple, rapid, highly sensitive and specific, which opened an alternative approach for the diagnosis of human angiostrongyliasis.

• Asian Pacific Journal of Cancer Prevention
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• 제17권12호
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• pp.5265-5272
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• 2016

Objective: The present study was conducted to evaluate invasive and noninvasive diagnostic methods for detection of Helicobacter pylori (H. pylori) in patients admitted with dyspeptic complaints and to compare sensitivities and specificities. Method: Sets of four gastric biopsy specimens were obtained from a total of 126 patients included in the study. The presence of H. pylori was determined by invasive tests including culture, rapid urease test, polymerase chain reaction (PCR) and histopathology. Among noninvasive tests, urea breath test, serological tests and enzyme-linked immunosorbent assay (ELISA) were performed. Results: H. pylori was isolated in 79 (62.7%) gastric biopsy cultures, whereas positivity was concluded for 105 (83.3%) patients by rapid urease test, for 106 (84.1%) by PCR, for 110 (87.3%) by histopathology, for 119 (94.4%) by urea breath test, and for 107 (84.9%) by ELISA. In the present study, the culture findings and histopathological examination findings were accepted as gold standard. According to the gold standard, urea breath test had the highest sensitivity (96.5%) and the lowest specificity (30%), whereas culture and histopathology had the highest specificities (100%). Conclusion: The use of PCR invasively with gastric biopsy samples yielded parallel results with the gold standard. PCR can be recommended for routine use in the diagnosis of H. pylori.

• Asian Pacific Journal of Cancer Prevention
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• 제17권6호
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• pp.2781-2785
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• 2016

Background: Matric metalloproteinase (MMP) 13 gene expression is increased in esophageal squamous cell carcinomas (ESCCs) and associated with increasing tumor invasion, lymph node involvement and decreased survival rates. Levels of the circulating enzyme may be elevated and used as a marker of tumor progression. In this study, clinical application of MMP-13 serum levels was evaluated for early detection, prediction of prognosis and survival time of ESCC patients. Materials and Methods: Serum levels of MMP13 were determined by ELISA in 66 ESCC patients prior of any treatment and 54 healthy controls for comparison with clinicopathological data through statistical analysis with Man Whitney U and Log-Rank tests. In addition, clinical value of MMP13 levels for diagnosis was evaluated by receiver operating characteristic (ROC) test. Results: The serum level of MMP-13 in patients (>250 pg/ml) was significantly higher than in the control group (<100 pg/ml) (p value=0.004). Also the results showed a significant correlation between MMP-13 serum levels with tumor stage (p value = 0.003), depth of tumor invasion (p value=0.008), involvement of lymph nodes (p value = 0.011), tumor size (p value = 0.018) and survival time. While there were no significant correlation with grade and location of tumors. ROC analysis showed that MMP-13 level is an accurate diagnostic marker especially to differentiate pre-invasive/ invasive lesions from normal controls (sensitivity and specificity: 100%). Conclusions: These findings indicate a potential clinical significance of serum MMP13 measurement for early detection and prognostic assessment in ESCC patients.

• Parasites, Hosts and Diseases
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• 제51권5호
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• pp.503-510
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• 2013

Toxoplasma gondii is an apicomplexan parasite with a broad host range of most warm-blooded mammals including humans, of which one-thirds of the human population has been infected worldwide which can cause congenital defects, abortion, and neonatal complications. Here, we developed a rapid diagnostic test (RDT) for T. gondii infection. Antigenic N-terminal half of the major surface antigen (SAG1) was linked with intrinsically unstructured domain (IUD) of dense granule protein 2 (GRA2). The recombinant GST-GRA2-SAG1A protein was successfully expressed and purified as 51 kDa of molecular weight. Furthermore, antigenicity and solubility of the rGST-GRA2-SAG1A protein were significantly increased. The overall specificity and sensitivity of GST-GRA2-SAG1A loaded RDT (TgRDT) were estimated as 100% and 97.1% by comparing with ELISA result which uses T. gondii whole cell lysates as the antigen. The TgRDT tested with Uganda people sera for field trial and showed 31.9% of seroprevalence against T. gondii antibody. The TgRDT is proved to be a kit for rapid and easy to use with high accuracy, which would be a suitable serodiagnostic tool for toxoplasmosis.

• Journal of Veterinary Science
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• 제21권4호
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• pp.68.1-68.8
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• 2020

A fluorescent microsphere-based immunochromatographic strip test (FICT) was developed for the rapid, sensitive, and quantitative detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies at the pen-side. The assay was based on the formation of a sandwich immune-complex (anti-pig IgG-PRRSV antibodies-NSP7/N), which was validated by a comparison with IDEXX-ELISA using 3325 clinical specimens. The diagnostic specificity, sensitivity, and accuracy of FICT were 97.28, 93.41, and 94.95%, respectively. FICT showed a good correlation with the virus neutralization assay. Overall, a promising pen-side diagnostic tool was developed for the rapid and quantitative detection of PRRSV antibodies within 15 min.

• 한국동물위생학회지
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• 제32권1호
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• pp.19-25
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• 2009

Porcine reproductive and respiratory syndrome (PRRS) virus is the etiological agent of diseases characterized by reproductive losses in sows and respiratory disorders in piglets. The PRRS virus is a small enveloped virus containing a positive-sense, single-stranded RNA genome. In the present study, ORF6 gene of Korean PRRS virus isolate, CNV, was cloned and expressed in baculovirus expression system. The ORF6 gene and expressed protein in the recombinant virus were confirmed by PCR/indirect fluorescence antibody (IFA) test and Western blotting, respectively. The recombinant protein with a molecular weight of approximately 24KDa was confirmed by Western blotting using His6 and PRRS virus-specific antiserum. Expressed ORF6 protein was applied for IFA to detect antibody against PRRS virus using field porcine sera. However, the sensitivity and specificity of developed IFA using expressed ORF6 protein were considerably low compared to those of commercial ELISA kit. This results suggest that IFA using expressed ORF6 protein could not be used as a diagnostic test for PRRS virus infection without further improvements.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.268-273
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• 2015

The fluorescent-antibody-to-membrane-antigen (FAMA) test is regarded as the "gold standard" to detect protective antibodies to varicella-zoster virus (VZV) because of its high sensitivity and specificity. Because the classic FAMA test uses an infectious virus for detection of antibodies to VZV, it is labor-intensive, and also requires special equipment for handling the virus. For this reason, we attempted to develop a simple and safe FAMA assay. Because VZV glycoprotein E (gE) is one of the major VZV glycoproteins, we used the gE protein for the FAMA test (gE FAMA). Here, we demonstrate that overexpression of gE in HEK293T cells can be used to measure antibodies in human serum, and that gE FAMA titers are closely correlated with gpEIA ELISA data. These results indicate that our gE FAMA test has the potential to measure antibodies to VZV.

• Parasites, Hosts and Diseases
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• 제24권2호
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• pp.171-176
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• 1986

In order to obtain the most specific and sensitive antigen from crude antigens of Fasciola hepatica for the immunodiagnosis of bovine fascioliasis by the enzyme linked immunosorbent assay (ELISA), phosphate buffered saline extract of F. hepatica was prepared. The crude extract was fractionated into 7 antigens using sephadex G-100 column chromatography. Seven fractionated antigens were applied to ELISA, precipitation test and intradermal test, respectively. Results obtained are as follows: 1. The specificity (95% confidence interval in negative sera of bovine fascioliasis; Mean+$2{\times}SD$ of absorbence) of the first (MW>150,000) and the second antigens(MW 120,000) were 93.7%, but those of others including crude antigen showed 100%. 2. The sensitivity(positive sera of bovine fascioliasis having higher values with compared to the criterion) of the first, the sixth (MW 16,000) and the seventh antigen (MW<5,000) were 91.6%, 87.5% and 0%, respectively, but those of others showed all 100%. 3. The absorbance by ELISA using the fifth antigen (MW 26,000) was 8. 43-folds higher in the positive sera than that in the negative sera. This could be used as one of the most specific antigens for the immunodiagnosis of bovine fascioliasis. 4. In Ouchterlony test, precipitin lines were not found in the sera naturally infected with F. hepatica, but some were found in the sera of rabbits immunized with the crude antigens. The numbers of precipitin lines in the sera of rabbits were different in the different fractionated antigens. They were 6 in the crude, 2 in the second and the third antigens, 1 in the fourth, the fifth and the sixth antigens and absent in the seventh antigen, respectively. 5. The wheal size for bovine infected with F. hepatica was $2.46{\pm}0.15cm$ in the intradermal test antigen(saline extract of F. hepatica) supplied by the Veterinary Research Institute, Rural Development Administration, Korea. The wheal size of the first, the second and the third antigens were larger than that. of intradermal test antigen, whereas those of the fouth, the fifth, the sixth and the seventh antigens showed smaller than that of the intradermal test antigen. The results suggest that the fifth antigen may be specific antigen for the immunodiagnosis of bovine fascioliasis.

• 핵의학기술
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• 제13권3호
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• pp.175-180
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• 2009

서론 : 갑상선 기능 항진을 보이는 환자를 대상으로 항갑상선제 치료후 재발유무를 예측하거나 정확한 그레이브스병을 진단하기 위해 임상에서는 갑상선자극호르몬 결합악제면 역글로블린을 측정하는데 지금까지 1세대 liquid phase (RIA Precipitate)를 사용하였다. 그 후 예민도가 높아진 다음 세대 측정법이 다양하게 개발되었으며 실제 임상적 유용성이 보고되었다. 따라서 세대별 갑상선자극호르몬 결합억제면역글로블린 측정법 임상적 유용성 검토해 보고자 한다. 실험재료 및 방법 : 2009년 1월에서 3월까지 본원 강남건강증진센터에 내원한 환자 중 TSH, FT4, Microsome Ab 측정값이 정상범위에 드는 30명 (남자 20명, 여자 10명)을 대조군으로 하였다. 마찬가지로 2009년 1월에서 3월까지 본원 내분비내과를 내원한 환자 중 TSH<$0.05\;{\mu}IU/mL$, FT4>1.80 ng/dL인 갑상선기능항진증 진단을 받은 58명 (남자14명, 여자 44명)을 실험군으로 하였다. 측정법은 각 세대별로 1세대에서 3세대까지 각각 2가지종류의 시약을 사용하였으며, 확진 검사로 TS-Ab 검사를 실시하였다. 결과 : 대조군을 대상으로 각 세대별 측정법의 특이도를 확인하였다. (Specificity=100%, n=30) 실험군에서 민감도는 1세대 RSR<%> (79.3%, n=58), 1세대 RSR (51.7%, n=58), 2세대 RSR-CT (93.1%, n=58), 2세대 BRAHMS-CT (98.3%, n=58), 3세대 ELISA (94.6%, n=56), 3세대 ECLIA (97.7%, n=58), TS-Ab<%> (93.5%, n=46)이었다. 결론 : 갑상선자극호르몬 결합억제면역글로블린 측정법의 세대별 검토 결과 TS-Ab(%)를 확진 검사로 기준했을 때 3세대 ECLIA에 대해 2세대 BRAHMS-CT 측정법이 가장 높은 상관성을 보였다($R^2$=0.923). 따라서 기존에 사용했던 1세대 측정법을 대신하여 2세대 측정법을 사용함으로써 보다 정확하고 신속한 결과보고에 유용할 것으로 판단된다.