• Journal of Oral Medicine and Pain
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• v.24 no.2
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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.

• The Korean Journal of Nuclear Medicine Technology
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• v.18 no.1
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• pp.153-157
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• 2014

Purpose: SLE (systemic lupus erythematosus) is an inflammatory autoimmune disease, characterized by various autoantibody. The detection of Anti double-stranded DNA (Anti-ds DNA) is important in the diagnostics of SLE, and include the American College of Rheumatology (ACR) diagnostic criteria for SLE. Also SLE disease activity and correlativity with the level Anti-ds DNA antibody have been reported and Anti-ds DNA antibody quantitative test is very useful for tracing before and after SLE treatment. When These Anti-ds DNA antibody test (Farr assay: $^{125}I$ labeled ds-DNA and bound Anti-ds DNA antibodies complex in serum is precipitated by ammonium sulfate and used to centrifugation, measured it) inhaled supernatant after centrifugation, a lipemic specimen does not facilitate the formation of precipitate and also occurs situation was inhaled with precipitate. To solve these problems, The Influence of the degree of lipemic specimen was evaluated. Materials and Methods: September 2012 to February 2013, We selected lipemic samples (n=81) of specimen commissioned by Anti-ds DNA antibody test. Lipemic samples were done pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) used a micro-centrifuge (Eppendorf Model 5415D). At the same time lipemic specimen and pre-treatment samples were performed Anti-ds DNA antibody test (Anti-ds DNA kit, Trinity Biotech, Ireland). Statistical analysis were analyzed Pearson's correlation coefficients and regression and paired t-test, and Difference (%). Results: Experimental group 1 (Lipemic Specimen Anti-ds DNA Ab concentration ${\leq}7IU/mL$) at y=0.368X+4.732, $R^2=0.023$, Pearson's correlation coefficient was 0.154, paired t-test (P=0.003), Difference (%) mean 65.7 and showed a statistically significant difference. Experimental group 2 (Lipemic Specimen Anti-ds DNA Ab concentration ${\geq}8IU/mL$) at y=0.983X+0.298, $R^2=0.994$, Pearson's correlation coefficient showed 0.997, paired t-test (P=0.181), Difference (%) mean -5.53 made no statistically significant difference. Conclusion: Lipemic sample of low Anti-ds DNA Ab concentration (2.5-7 IU/mL) and the result is obtained pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) were made a significant difference statistically. Anti-ds DNA is one of the primary auto-antibodies present in patients with SLE, and remain an important diagnostic test for SLE. Therefore, we recommend preprocessing (high-speed centrifugation: 14,000 rpm 5 mins) in order to exclude the influence of lipemic specimen.

• Korean Journal of Veterinary Research
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• v.37 no.2
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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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• v.12 no.2
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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.

• Biomedical Science Letters
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• v.11 no.1
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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.

• Korean Journal of Veterinary Service
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• v.39 no.2
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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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• v.41 no.4
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• pp.354-362
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• 1994

Objectives and Methods : The presence of aneuploidy or high proliferative activity in cytologic specimens is considered as complementary for the diagnosis of malignancy. To evaluate the diagnostic usefulness of DNA ploidy and cell cycle analysis in lung cancer, we compared the diagnostic yielding rates of DNA ploidy test by brushing specimens using flow cytometry with bronchoscopic forceps biopsy and brushing cytology. Results : Of the seventy-six cases, 55 cases proved to have malignant diseases(squamous cell cancer: 27, adenocarcinoma: 7, large cell cancer: 1, undifferentiated: 4 and small cell cancer: 16). The incidence of aneuploidy in lung cancer patients was 32.7%(18/55), as opposed to no cases in benign disease. And the proportion of high proliferative activity(S+G2M>22%) in lung cancer patients was 42.9%(15/35), but none in benign diseases. In fifty-six of 75 cases(74.7%), cytology of brushing specimens and DNA analysis(either aneuploidy or high proliferative activity vs. diploidy and low proliferative activity) were in concordance. The sensitivity with only brushing cytology was 41.8%(23/55), but with the addition of DNA analysis, it was increased to 56.4%(31/55), without decreasing the specificity(100%). And there was a case whose clue for malignancy was absent except aneuploidy, and he was confirmed to have squamous cell cancer following open thoracotomy. There were no differences in the frequency of aneuploidy or high proliferative activity between histologic subtypes of bronchogenic malignancy. Conclusions : The diagnostic detection rate of lung cancer was improved with the addition of DNA ploidy and cell cycle analysis, and the presence of aneuploidy or high proliferative activity was a relatively specific indicator of malignant disease. It would be useful to test DNA ploidy and cell cycle analysis with brushing specimen for the diagnosis of bronchogenic malignancy particularly in patients whose biopsy specimen could not be obtainable.

• Korean Journal of Veterinary Research
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• v.39 no.6
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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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• v.35 no.6
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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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• v.53 no.3
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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.