• Journal of Oral Medicine and Pain
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• 제24권2호
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• pp.181-187
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• 1999

The most important progress in diagnostic sciences is the increased sensitivity and specificity in diagnostic procedures due to the development of newer micromethodologies and increasing availability of immunological and molecular biological reagents. The outcome of researches in this field has already provided DNA probes and antibodies which can be used for diagnosing various kinds of diseases including inherited ones. This development can be also applied to diagnose diseases in oral and maxillofacial regions. Technological advances have yielded highly sensitive test methodologies so that low analyte concentration and small sample volume are no longer limiting factors. Therefore, saliva can be useful test fluid for an array of analytes. Salivary constituents of diagnostic significance include steroid hormones, antibodies, drugs, and tumor markers. Of the proteins present in saliva, viral-specific immunoglobulins are of the greatest diagnostic interest. The development of conjugates and antigens by recombinant DNA technique and peptide synthesis is necessary for clinical application. Several kits developed for the purpose of blood testing should be modified to permit their application to saliva. The final practical outcome of researches in diagnostic sciences will be various diagnostic agents which can be used for detection of bacteria and viruses, screening and diagnosis of diseases, genetic screening for forensic individual identification. For these purposes, collaboration researches and development between institutions and companies are essential.

• 핵의학기술
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• 제18권1호
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• pp.153-157
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• 2014

Anti double-stranded DNA (Anti-ds DNA)항체의 검출은 SLE의 진단에 중요하며 American College of Rheumatologists의 SLE 진단기준에 포함되어 있다. 또한 SLE 질병의 활성도와 Anti-ds DNA 항체 수준의 상관성이 보고되어 있으며 Anti-ds DNA 항체 정량검사는 SLE의 치료 전, 후 추적에 매우 유용하다. Anti-ds DNA 항체의 검출을 위한 검사 방법으로 방사면역측정법(radioimmunoassay, RIA)을 이용한 Farr assay, Crithidia luciliae를 이용한 면역형광 측정법(immunofluorescence Test, CLIFT), 효소면역측정법(enzyme-linked immunosorbent assay, ELISA), 화학발광면역 측정법(chemiluminescence immunoassay, CLIA)이 있다. 본원에서 방사면역측정법(radioimmunoassay, RIA)으로 Anti-ds DNA 항체 검사 과정에서 lipemic한 검체의 경우 침전물의 형성이 원활하지 않거나 침전물도 같이 흡입되는 상황이 발생 하였고, 이러한 문제점을 해결하기 위해 lipemic 검체가 분석 결과에 미치는 영향 정도를 평가하였다. 2012년 9월부터 2013년 2월까지 Anti-ds DNA 항체 검사가 의뢰된 검체 중 lipemic한 검체(n=81)를 선택하여 마이크로 원심분리기로 전처리(고속 원심분리: 14,000 rpm 5 mins)한 후 동시에 Anti-ds DNA 항체 검사(Anti-ds DNA kit, Trinity Biotech, Ireland)를 시행하였다. 실험군 1 (lipemic 검체의 Anti-ds DNA 항체 농도 ${\leq}7IU/mL$)에서 y=0.3636x+4.7322, $R^2=0.0238$, Pearson 상관계수는 0.154, paired t-test (P=0.003), Difference (%) mean 65.7의 결과를 얻었으며 통계적으로 유의한 차이를 보였다. 그러나 실험군 2 (lipemic 검체의 Anti-ds DNA 항체 농도 ${\geq}8IU/mL$)에서 y=0.9837x+0.2982, $R^2=0.994$, Pearson 상관계수는 0.997, paired t-test (P=0.181), Difference (%) mean -5.53을 보였고 통계적으로 유의한 차이가 없음을 확인할 수 있었다. 임상에서 SLE (systemic lupus erythematosus)의 진단에 중요한 역할을 하는 Anti-ds DNA 항체 검사는 lipemic 검체의 영향을 배제하기 위해서 반드시 전처리(고속 원심분리: 14,000 rpm 5 mins) 과정을 통해 혈청 내 지질 성분을 제거한 후 검사를 시행하여야 한다.

• 대한수의학회지
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• 제37권2호
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• pp.349-358
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• 1997

Mycobacterium paratuberculosis is the etiologic agent of Johne's disease, a chronic inflammatory bowel syndrome in ruminants. The attempts to control or eradicate the disease were severely hampered by the inadequacies of present diagnostic methods. The first purpose of this study was to detect Johne's disease out of 577 cows in the province of Kyunggi, Chungchong, Gangweon and the second purpose was to compare the results of non-absorbed ELISA, absorbed ELISA, PCR, and conventional culture methods. The third purpose was to increase diagnostic specificity, accuracy and rapidity. When non-absorbed ELISA test was conducted with Mycobacterium paratuberculosis antigen, the prevalence of positive was 10.9%. To increase diagnostic specificity, absorbed ELISA test with Mycobacterium phlei was used. In this test, the positive prevalence was 1.7%. For the specific detection of Mycobacterium paratuberculosis, PCR was applied to bacterial culture obtained from fecal samples of cattle. The DNA sequences derived from IS900 were used to prepare DNA primers for detection and identification of Mycobacterium paratuberculosis by PCR. PCR for M paratuberculosis isolated from fecal cultures amplified specific target DNA. PCR was much more rapid than that obtained by conventional culture technique in diagnosis of Johne's disease.

• Journal of Genetic Medicine
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• 제12권2호
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• pp.66-71
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• 2015

Down syndrome screening with cell-free DNA (cfDNA) in the maternal plasma has recently received much attention in the prenatal diagnostic field. Indeed, a large amount of evidence has already accumulated to show that screening tests with cfDNA are more sensitive and specific than conventional maternal serum and/or ultrasound screening. Globally, more than 1,000,000 of these noninvasive prenatal tests (NIPTs) have been performed to date. There are several different methods for NIPTs that are currently commercially available, including shotgun massively parallel sequencing, targeted massively parallel sequencing, and single nucleotide polymorphism (SNP)-based methods. All of these methods have their own advantages and disadvantages. In this review, I will focus specifically on the SNP-based NIPT.

• 대한의생명과학회지
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• 제11권1호
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• pp.51-56
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• 2005

The detection of high-risk human papilloma virus (HPV) allows us to predict the presence and future development of cervical intraepitheliallesion. In this study, we compared Hybrid Capture II and DNA chip methods for detection of HPV in cervical swab samples. And we evaluated the clinical efficacy and diagnostic performance of HPV DNA chip and Hybrid Capture II for detecting HPV in cervical neoplastic lesions. Seventy four patients were classified into three groups according to their histologic diagnosis: Group I (nonspecific chronic cervicitis), Group II (low-grade squamous intraepithelial lesion (SIL); koilocytosis, and mild dysplasia), and Group III (high-grade SIL;, moderate, severe dysplasia and in situ carcinoma). Cytologic diagnosis were based on the Bethesda System. Hybrid Capture II and DNA chip methods were performed to detect HPV. In 41 of the 74 cervical samples $(55.4\%)$, HPV DNAs were detected by Hybrid Capture II. In Group III, HPV-positive cases were detected in 15 $(20.3\%)$ of 74 patients by Hybrid Capture II. 25 patients with ASCUS cytology were histopathologically examined: 9 cases $(36\%)$ were Group II. In 18 patients with low-grade SIL cytology, 13 cases $(72.2\%)$ were Group II and 3 cases $(16.7\%)$ were Group III. 12 cases $(92.3\%)$ were Group ill of 13 patients with high-grade SIL cytology. The sensitivity of each test was $82\%$ in Hybrid Capture II and $53.9\%$ in DNA chip test. And the specificity was $74.3\%,\;85.7\%$ in Hybrid Capture II and DNA chip. In conclusion, Hybrid Capture II test is more sensitive than DNA chip in detecting women with cervical neoplastic lesions. Especially, in diagnosing of ASCUS, Hybrid Capture II test is more sensitive. Therefore, Hybrid Capture II test for cancer-associated HPV DNA is a viable option in the management of women with ASCUS.

• 한국동물위생학회지
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• 제39권2호
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• pp.107-116
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• 2016

In the study, we developed and evaluated a uracil N-glycosylase (UNG)-supplemented single-tube nested reverse transcription-polymerase chain reaction (UsnRT-PCR) assay that can carried out first-round RT-PCR and second-round nested PCR in a reaction tube without reaction tube opening and can simultaneously detect EU- and NA-PRRSV. The UsnRT-PCR confirmed to have a preventing ability of mis-amplification by contamination of pre-amplified PRRSV DNA from previous UsnRT-PCR. Primer specificities were evaluated with RNAs extracted from 8 viral strains and our results revealed that the primers had a high specificity for both genotypes of PRRSV. The sensitivity of the UsnRT-PCR was 0.1 $TCID_{50}$/0.1 mL for EU- or NA-PRRSV, respectively, which is comparable to that of previously reported real time RT-PCR (RRT-PCR). Clinical evaluation on 110 field samples (60 sera and 50 lung tissues) by the UsnRT-PCR and the RRT-PCR showed that detection rates of the UsnRT-PCR was 70% (77/110), and was relatively higher than that of the RRT-PCR (69.1%, 76/110). The percent positive or negative agreement of the UsnRT-PCR compared to RRT-PCR was 96.1% (73/76) or 90.9% (30/33), showing that the test results of both assays may be different for some clinical samples. Therefore, it is recommend that diagnostic laboratory workers use the two diagnostic assays for the correct diagnosis for the relevant samples in the swine disease diagnostic laboratories. In conclusion, the UsnRT-PCR assay can be applied for the rapid, and reliable diagnosis of PRRSV without concerns about preamplified DNA carryover contamination that can occurred in PCR process in the swine disease diagnostic laboratories.

• Tuberculosis and Respiratory Diseases
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• 제41권4호
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• pp.354-362
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• 1994

배경 및 방법 : 폐암의 진단 방법에 있어서 객담, 흉수, 기관지 세척액 등에서의 DNA의 aneuploid나 고증식력등의 소견은 폐암의 진단에 보조적으로 의의가 있는 것으로 보고되고 있다. 저자들은 기관지 내시경 검사 중에 조직 생검, 솔질표본의 세포학적 검사와 더불어 유세포계산법을 이용한 솔질 표본의 DNA 배수성 검사를 함께 시행하여, 폐암의 진단율을 높일 수 있는지의 여부를 검토하고자 하였다. 결과: 1) 대상환자 76예중 폐암으로 확진되었던 55예에서는 diploid 37예, aneuploidy 18예(32.7%)이었으나, 양성 질환으로 확인된 21예에서는 모두 diploid 이었고, 세포주기 분석이 기능하였던 48예중 폐암은 35예 이었고 이들중 42.9%(15/35)에서 고증식력을 보였으나, 양성질환 13예에서는 고증식력을 보인 경우가 없었다(p<0.05). 2) DNA분석 소견(aneuploidy나 고증식력을 양성으로 하였을 때)과 세포진검사와의 일치율은 전체 75예중 56예로 74.7% 였다. 3) 폐암 환자에서 세포진검사 민감도는 41.8%(23/55)이었는데, 세포진검사 음성이지만 DNA 검사에서 양성(aneuploidy 혹은 high proliferative activity)을 보인 경우를 부가하였을 때, 민감도는 56.4%(31/55)로 증가하였고(p<0.05), 음성예측도는 38.2%, 특이도는 100% 였다. 4) 1예에서는 기관지내시경을 이용한 조직, 세포진검사, 경피적 폐침흡인 등으로는 진단을 내릴수 없었으나 솔질표본을 이용한 DNA ploidy검사에서 aneuploid로 나타났고, 후에 수술로써 편평상피폐암으로 확진되었다. 5) 폐암 환자중 세포 형태에 따른 DNA Ploidy와 증식력에는 유의한 차이가 없었다. 결론 : 기관지 솔질 표본에서 aneuploid나 고증식력 소견이 폐암을 시사하는 것으로 해석하였을 때, 세포진검사에 부가하여 DNA 배수성 측정과 세포주기 분석을 함께 함으로써 폐암 진단의 예민도를 높일수 있었고, 비교적 특이도가 높은 것으로 사료되었으며, 특히 조직을 얻기 어려운 경우들에서 DNA 분석의 진단적 의의에 대한 계속적인 연구가 필요할 것으로 사료되었다.

• 대한수의학회지
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• 제39권6호
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• pp.1151-1160
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• 1999

This study was conducted to detect the toxoplasma-specific DNA in peripheral blood collected from cats experimentally infected with Toxoplasma gondii (RH strain) and from domiciled cats by B1 gene-base polymerise chain reaction(PCR). The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with parasite detection by mouse inoculation(MI). And also, latex agglutination test(LAT) and indirect fluorescent antibody test(IFAT) were conducted to detect the fluctuation of serum antibodies compared with the detection of toxoplasma by PCR and MI. Toxoplasma B1 gene PCR was shown consistently high sensitivity and the results obtained by PCR agreed completely with those from MI. All blood samples collected before infection with T gondii gave negative results by PCR and MI. Also, toxoplasma Bl gene PCR was not cross reaction with Neospora caninum DNA and normal cat leucocyte as controls. The toxoplasma-specific DNA was detected by PCR in blood of 5 cats experimentally infected with T gondii 6 days after infection and the detection of this specific-DNA was long lasted in blood for 64 days after infection. The detection of toxoplasma-specific DNA by PCR could be identified as few as 10 tachyzoites and the isolation of T gondii by MI could be isolated as few as 1 tachyzoite from tenfold serial dilution of T gondii with normal cat blood, respectively. In healthy domiciled cats, the toxoplasma-specific DNA and the parasite were detected and isolated in blood from 3 of 56(5.3%) cats by both PCR and MI, respectively. In the results of antibody test from the total 56 heads of healthy domiciled cats, the positive rates are 15(26.7%) by LAT and 19(33.9%) by IFAT. These results suggest that PCR detection of toxoplasma can be applied as a sensitive and specific diagnostic and research tool.

• The Plant Pathology Journal
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• 제35권6호
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• pp.635-643
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• 2019

To detect Xanthomonas arboricola pv. pruni, a loopmediated isothermal amplification (LAMP) detection method were developed. The LAMP assay was designed to test crude plant tissue without pre-extraction, or heating incubation, and without advanced analysis equipment. The LAMP primers were designed by targeting an ABC transporter ATP-binding protein, this primer set was tested using the genomic DNA of Xanthomonas and non-Xanthomonas strains, and a ladder product was generated from the genomic DNA of X. arboricola pv. pruni strain but not from 12 other Xanthomonas species strains and 6 strains of other genera. The LAMP conditions were checked with the healthy leaves of 31 peach varieties, and no reaction was detected using either the peach leaves or the peach DNA as a template. Furthermore, the high diagnostic accuracy of the LAMP method was confirmed with 13 X. arboricola pv. pruni strains isolated from various regions in Korea, with all samples exhibiting a positive reaction in LAMP assays. In particular, the LAMP method successfully detected the pathogen in diseased peach leaves and fruit in the field, and the LAMP conditions were proven to be a reliable diagnostic method for the specific detection and identification of X. arboricola pv. pruni in peach orchards.

• Clinical and Experimental Pediatrics
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• 제53권3호
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• pp.273-285
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• 2010

Completion of the human genome project has allowed a deeper understanding of molecular pathophysiology and has provided invaluable genomic information for the diagnosis of genetic disorders. Advent of new technologies has lead to an explosion in genetic testing. However, this overwhelming stream of genetic information often misleads physicians and patients into a misguided faith in the power of genetic testing. Moreover, genetic testing raises a number of ethical, legal, and social issues. Diagnostic genetic tests can be divided into three primary but overlapping categories: cytogenetic studies (including routine karyotyping, high-resolution karyotyping, and fluorescent in situ hybridization studies), biochemical tests, and DNA-based diagnostic tests. DNA-based testing has grown rapidly over the past decade and includes preandpostnatal testing for the diagnosis of genetic diseases, testing for carriers of genetic diseases, genetic testing for susceptibility to common non-genetic diseases, and screening for common genetic diseases in a particular population. Theoretically, once a gene's structure, function, and association with a disease are well established, the clinical application of genetic testing should be feasible. However, for routine applications in a clinical setting, such tests must satisfy a number of criteria. These criteria include an acceptable degree of clinical and analytical validity, support of a quality assurance program, possibility of modifying the course of the diagnosed disease with treatment, inclusion of pre-and postnatal genetic counseling, and determination of whether the proposed test satisfies cost-benefit criteria and should replace or complement traditional tests. In the near future, the application of genetic testing to common diseases is expected to expand and will likely be extended to include individual pharmacogenetic assessments.