• Asian-Australasian Journal of Animal Sciences
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• 제16권7호
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• pp.1046-1048
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• 2003

A polymerase chain reaction (PCR) assay was developed to differentiate buffalo meat from the meat of Ceylon spotted deer (Axis axis ceylonensis), Ceylon sambhur (Cervus unicolor unicolor), cattle (Bovine), goat (Caprine), pig (Porcine), and sheep (Ovine). A set of primers were designed according to the sequence of the mitochondrial cytochrome b gene of bubalus bubalis and by PCR amplification a band of approximately 242 bp band was observed with buffalo DNA. These primers did not cross-react with DNA of other animal species tested in the study under the specified reaction conditions. A band of 649 bp was observed for all animal species tested when DNA was amplified with the universal primers indicating the presence of mitochondrial DNA in the samples. The technique was sensitive enough to identify rotten (10 days post slaughter), dried and cooked buffalo meat. The absence of a cross reaction with human DNA using the buffalo specific primers eliminates possible false positive reactions.

• 대한바이러스학회지
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• 제30권2호
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• pp.151-159
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• 2000

Hepatitis C virus (HCV) is one of the important human pathogen that can cause acute and chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. Recently, the third generation radiation immuno assay (RIA) method has been developed as a very sensitive test to detect anti-HCV antibody. However, false positive is the problem with RIA test. To solve this the RIA results were compared to those of 5-antigen recombinant immunoblot assay (5-RIBA) and reverse transcription-polymerase chain reaction (RT-PCR). Among 12,767 serum samples tested from clinic visitors, total 275 (2.2%) samples were antibody positive by RIA. RIBA was performed with 148 RIA positives cases but among them was shown eighty five was antibody positive and sixty three (42.6%) was negative result. However, nested RT-PCR test was shown also carried out with 43 positive, 6 intermediates and 25 negatives of RIBA. As a result of the nested RT-PCR results, HCV antigen were detected in RIBA positive, 33.3% (2/6) RIBA intermediate and 12% (3/25). Clinical syndrome of all 148 patients as a with chronic active hepatitis (46.0%), cirrhosis (18.9%), hepatocellular carcinoma (8.1%) and others (27.0%) and they were positive in reaction by RIA test. But RIBA positive patients with 34.9% of chronic active hepatitis, 18.6% of cirrhosis, 4.6% of hepatocellular carcinoma and 41.9% of others were detected to be positive case by nested RT-PCR.

• 대한임상검사과학회지
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• 제42권1호
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• pp.38-45
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• 2010

The aim of this study was to assess the Clinical Usefulness of Helicobacter pylori Stool Antigen (HpSA) immunochromatographic assay for the diagnosis of H. pylori infection. In this study, we had compared HpSA-immunochromatographic assay with CLO test and UBT test. From a total of 140 patients (M:F=88:52) with upper endoscopy, biopsy specimens were obtained for CLO test. Stool specimens was collected from all patients and tested using a HpSA-immunochromatic assay. H. pylori infection status was defined as infected if the results of both CLO test and UBT test were positive. CLO test and UBT test findings showed that 92 patients were H. pylori positive and 48 patients were H. pylori negative. According to this definition, the sensitivity, specificity, and positive or negative predictive value (PPV, NPV) of HpSA-immunochromatographic assay were 97.8%, 100%, 100%, and 96%, respectively. Cross reactivity test of HpSA-immunochromatographic assay were performed with 10 enteric bacteria strains in fecal habitat, and there were no false positive reaction. We evaluated the usefulness of HpSA assay for eradication therapy with 10 of 92 H. pylori positive patients, positive results of them at pre-eradication therapy were converted to negative at post-eradication. The HpSA-immunochromatographic assay is a highly sensitive and specific non-invasive diagnostic method for detection of H. pylori infection, a useful diagnostic method for H. pylori in post eradication stage.

• Journal of Microbiology and Biotechnology
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• 제30권3호
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• pp.459-468
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• 2020

This study established a new polymerase spiral reaction (PSR) that combines with reverse transcription reactions for HCV detection targeting 5'UTR gene. To avoid cross-contamination of aerosols, an isothermal amplification tube (IAT), as a separate containment control, was used to judge the result. After optimizing the RT-PSR reaction system, its effectiveness and specificity were tested against 15 different virus strains which included 8 that were HCV positive and 7 as non-HCV controls. The results showed that the RT-PSR assay effectively detected all 8 HCV strains, and no false positives were found among the 7 non-HCV strains. The detection limit of our RT-PSR assay is comparable to the real-time RT-PCR, but is more sensitive than the RT-LAMP. The established RT-PSR assay was further evaluated for detection of HCV in clinical blood samples, and the resulting 80.25% detection rate demonstrated better or similar effectiveness compared to the RT-LAMP (79.63%) and real-time RT-PCR (80.25%). Overall, the results showed that the RT-PSR assay offers high specificity and sensitivity for HCV detection with great potential for screening HCV in clinical blood samples.

• Pediatric Infection and Vaccine
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• 제7권1호
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• pp.113-119
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• 2000

Purpose : Rotavirus is a most common etiologic agent of pediatric gastroenteritis. The standard method to diagnose rotavirus infection was the detection of viral particles in specimens through electron microscopy. But it was complex. Enzyme immunoassay and latex agglutinin are preferred because they are relatively handy, inexpensive and take a short time, in comparison with electron microscopy. However, several reports have shown that the use of ELISA to diagnose rotavirus infection in neonates can result in false positive reactions. The main purpose of this study is to compare ELISA and RT-PCR in the diagnosis of neonatal rotavirus infection. Methods : Data presented in this study were obtained form 123 newborn babies in the nursery of the Fatima Hospital, Masan, Korea, form Jury to December, 1997. We obtained two samples of stool from each of the newborn babies and then performed the Rotazyme test and the RT-PCR. In the Rotazyme test, the results were interpreted according to visual findings. The samples were used for the RT-PCR test after at stock $-30^{\circ}C$ to identify rotavirus group A. The result of the two tests were compared. Results : The informations are divided into 73 males and females. Out of the total informations 15 were transferred from other hospitals. Their average gestational age was $38.5{\pm}1.6$ weeks. The average birth weight was $3134.8{\pm}539gm$. In the Rotazyme test, 75 samples turned out to be positive. Out of them, 55 samples(75.3%) were positive and 18 samples(24.7%) were negative in the RT-PCR. On the other hand, in the Rotazyme test, 50 samples turned out be negative. Out of them, 27 samples(54%) were positive and 23 samples(46%) were negative in the RT-PCR. Conclusion : Rotavirus infection in uncommon in neonates. The diagnosis based on visual findings using Rotazyme test has a disadvantage in the sense that it can result in false positive reactions and false negative reactions in the diagnosis of neonatal rotavirus infection.

• 한국환경보건학회지
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• 제45권5호
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• pp.511-519
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• 2019

Objectives: The standard method for the enumeration of environmental Legionella is culturing, which has several disadvantages, including long incubation and poor sensitivity. The purpose of this study is to demonstrate the usefulness of real-time PCR and to improve the standard method. Methods: In 200 environmental water samples, a real-time PCR and culture were conducted to detect and quantify Legionella. Using with the results of the survey, we compared the real-time PCR with the culture. Results: Each real-time PCR assay had 100% specificity and excellent sensitivity (5 GU/reaction). In the culture, 36 samples were positive and 164 samples were negative. Based on the results of the culture, real-time PCR showed a high negative predictive value of 99%, 35 samples were true positive, 105 samples were true negative, 59 samples were false positive and one sample was a false negative. Quantitative analysis of the two methods indicated a weak linear correlation ($r^2=0.29$, $r^2=0.61$, respectively). Conclusions: Although it is difficult to directly apply quantitative analysis results of real-time PCR in the enumeration of environmental Legionella, it can be used as a complementary means of culturing to rapidly screen negative samples and to improve the accuracy of diagnosis.

• Asian-Australasian Journal of Animal Sciences
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• 제33권10호
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• pp.1683-1690
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• 2020

Objective: The rapid and reliable detection of the African swine fever virus (ASFV) plays an important role in emergency control and preventive measures of ASF. Some methods have been recommended by FAO/OIE to detect ASFV in clinical samples, including realtime polymerase chain reaction (PCR). However, mismatches in primer and probe binding regions may cause a false-negative result. Here, a slight modification in probe sequence has been conducted to improve the qualification of real-time PCR based on World Organization for Animal Health (OIE) protocol for accurate detection of ASFV in field samples in Vietnam. Methods: Seven positive confirmed samples (four samples have no mismatch, and three samples contained one mutation in probe binding sites) were used to establish novel real-time PCR with slightly modified probe (Y = C or T) in comparison with original probe recommended by OIE. Results: Both real-time PCRs using the OIE-recommended probe and novel modified probe can detect ASFV in clinical samples without mismatch in probe binding site. A high correlation of cycle quantification (Cq) values was observed in which Cq values obtained from both probes arranged from 22 to 25, suggesting that modified probe sequence does not impede the qualification of real-time PCR to detect ASFV in clinical samples. However, the samples with one mutation in probe binding sites were ASFV negative with OIE recommended probe but positive with our modified probe (Cq value ranked between 33.12-35.78). Conclusion: We demonstrated for the first time that a mismatch in probe binding regions caused a false negative result by OIE recommended real-time PCR, and a slightly modified probe is required to enhance the sensitivity and obtain an ASF accurate diagnosis in field samples in Vietnam.

• 대한정형외과 초음파학회지
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• 제3권2호
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• pp.54-58
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• 2010

목적: 만성적으로 경추 동통을 호소하는 환자의 통증의 근원은 대부분 후관절인데, 증상을 일으키는 관절의 정확한 위치 확인의 어려움과 합병증 및 위약효과의 가능성으로 인해 임상에서의 적용에 의구심을 가지게 된다. 본 연구에서는 만성 경추 동통을 호소하는 환자에서 후관절 동통의 유병율을 평가하고 진단 및 치료적 목적의 초음파를 이용한 후관절 차단술의 유용성에 대해 보고한다. 대상 및 방법: 만성적이면서 비특이적인 경추 동통을 6개월 이상 호소하는 환자를 대상으로 하였다. 방사통을 동반한 추간판 탈출 관련 통증을 호소하는 환자는 그 대상에서 제외되었으며, 물리 치료나 척추 교정 요법, 약물 치료 등의 보존적 치료 요법에 실패한 환자 160명을 대상으로 하였다. 경추 동통에 대한 진단적 후관절 차단술은 1% 리도카인(lidocaine)을 사용하여 시행하였으며, 리도카인에 양성 반응을 보인 환자에 대해서는 2주 뒤 0.25% 부피바카인(bupivacaine)을 사용하였으며, 후관절 차단술 시행 이후 경추 운동시 75% 이상의 통증 감소를 보일 경우를 양성 반응으로 고려하였다. 결과: 만성 경추 동통을 호소하는 환자 160명 가운데 리도카인을 이용한 후관절 차단술에 96명, 60% 양성반응을 보였으며, 리도카인에 음성반응을 보인 환자 64명 가운데 부피바카인에 48명, 75% 양성반응을 보였다. 리도카인에 양성반응을 보였으나 부피바카인에 음성반응을 보인 가양성 환자는 48명, 50%에 해당하였다. 총 160명의 환자들의 시술 전 VAS는 평균 8.5이었으며, 2주 추시시 2.7 (p=0.001), 4주 추시시 3.6 (p=0.001)로 감소하였다. 초기 시술로 증상의 호전이 만족스럽지 않았던 8명과 경과 관찰 도중에 악화되었던 3명은 4주째 후관절 차단술을 추가적으로 실시하여 통증의 완화를 보였다. 결론: 만성 경추 동통을 호소하는 환자에 대해 초음파를 이용한 단일 차단술은 가양성 반응이 높아 진단적 가치가 상대적으로 낮으며, 후관절에 시행한 두 번의 차단술은 만성 경추 동통 환자의 중재적 통증 조절에 유용한 방법으로 사료된다.

• 대한미생물학회지
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• 제22권4호
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• pp.445-451
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• 1987

Coagglutination method is widely used for the diagnosis of Salmonella infection. This test, however, has a disadvantage of false positive reaction due to the coagglutination of staphylococci with non-specific immune complexes or anti-staphylococci antibody in serum. Salmonell O antigen was detected by enzyme immunoassay with protein A-bearing Staphylococcus aureus as in the solid phase. Horse radish peroxidase was labeled to IgG specific against Salmonella O antigen. This enzyme immunoassay was much more sensitive than conventional coagglutination method without false poitive agglutination. To improve the sensitivity for detection of Salmonella O antigen in samples, we tried to determine the optimal concentration of normal IgG that inhibits non-specific binding of horse radish peroxidase labeled IgG to staphylococci, and to establish the optimal condition of reaction between antigen-antibody complex and staphylococci. Non-specific binding of horse radish peroxidase labeled specific IgG to staphylococci was almost blocked when the enzyme labeled IgG was 500-fold diluted with phosphate buffered saline containing 2mg/ml of normal IgG. When staphylococci coated with antibody to Salmonella O antigen were mixed with antigen-antibody complex and then incubated for 1 hour at room temperature, the minimal detectable concentration of Salmonella O antigen was 1ng/ml. The sensitivity of enzyme immunoassay was 100-fold greater than a conventional coagglutination method. This enzyme immunoassay could be expected as an improved method for detection of other infectious agents.

• 대한수의학회지
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• 제43권3호
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• pp.471-476
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• 2003

Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by ovine herpesvirus 2 (OvHV-2). OvHV-2 is a gamma herpesvirus, which induces frequent latent infection and often difficult to detect its antigens and even specific nucleic acids because of its low viral copies in the infected tissues. Histopathology, serology and polymerase chain reaction (PCR) were compared for the diagnosis of MCF using 10 bison infected with OvHV-2. Histopathological diagnosis was performed using the criteria which was based upon the pathognomic lesions. Serological diagnosis was conducted using its serum with competitive ELISA for the detection of antibodies of OvHV-2. Also, the nest PCR was performed with peripheral blood leukocytes for the detection of OvHV-2-specific DNAs. Primers 556 and 775 were used for the primary amplification, and primers 556 and 555 were used for the secondary amplification. As the results, positive cases were 6 by histopahology, 9 by serology and 10 by PCR. As comparing with other diagnostic methods, PCR was found to be more sensitive than histopathology and serology. The recent development of molecular diagnostic assays has provided powerful tools for investigating how viruses survive in nature. Development of PCR specific for viruses has dramatically improved the accuracy of diagnosis of viruses in clinically infected animals. Furthermore, amplification of viral genomic material by nest PCR represents the most sensitive method for the detection of viruses and might be detected successfully even though very low viral DNA copies. So, it could be used as the first choice for the detection of viral DNAs with low copies such as the status of latent infection. However, it has also some limitation of application like as false negative results by PCR inhibitors and false positive results by contamination. The results of this study suggest that the use of molecular biological methods like PCR may increase the accuracy for the diagnosis of infectious diseases. However, in diagnostic laboratory, it is recommended that PCR assay must be conducted with other diagnostic methods for more reliable diagnosis.