• Title/Summary/Keyword: PCR diagnosis

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Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Korea

  • Seo, Min-Goo;Ouh, In-Ohk;Choi, Eunsang;Kwon, Oh-Deog;Kwak, Dongmi
    • Parasites, Hosts and Diseases
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    • v.56 no.6
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    • pp.559-565
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    • 2018
  • The identification and characterization of pathogenic and zoonotic tick-borne diseases like granulocytic anaplasmosis are essential for developing effective control programs. The differential diagnosis of pathogenic Anaplasma phagocytophilum and non-pathogenic A. phagocytophilum-like Anaplasma spp. is important for implementing effective treatment from control programs. The objective of the present study was to investigate the prevalence of Anaplasma spp. in horses in Korea by nucleotide sequencing and restriction enzyme fragment length polymorphism assay. Of the 627 horses included in the study, only 1 (0.2%) was infected with A. phagocytophilum. Co-infection with A. phagocytophilumlike Anaplasma spp. was not detected in the study. The 16S rRNA sequence of A. phagocytophilum was similar (99.5-100%) to A. phagocytophilum 16S rRNA isolated from horses in other countries. PCR adapted to amplify A. phagocytophilum groEL and msp2 genes failed to generate amplicons, suggesting genetic diversity in these genes. This study is the first molecular detection of A. phagocytophilum in horses in Korea. Human granulocytic anaplasmosis and animal infection of A. phagocytophilum have been reported in Korea recently. Because of vector tick distribution, global warming, and the increase of the horse industry, horses should be considered as a potential reservoir for A. phagocytophilum, and cross infectivity should be evaluated even though a low prevalence of infection was detected in this study. Furthermore, continuous surveillance and effective control measures for A. phagocytophilum should be established to prevent disease distribution and possible transmission to humans.

Molecular Identification of Anisakis Larvae Extracted by Gastrointestinal Endoscopy from Health Check-up Patients in Korea

  • Song, Hyemi;Jung, Bong-Kwang;Cho, Jaeeun;Chang, Taehee;Huh, Sun;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.57 no.2
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    • pp.207-211
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    • 2019
  • Anisakiasis is a zoonotic disease induced by anisakid nematodes, and endoscopic inspection is used for a diagnosis or remedy for it. Anisakis simplex, Anisakis physeteris, and Pseudoterranova decipiens had been reported to be the major species causing human infections, particularly, in Japan. However, in Korea, recent studies strongly suggested that Anisakis pegreffii is the major species of human infections. To support this suggestion, we collected anisakid larvae (n=20) from 20 human patients who were undergone gastrointestinal endoscopy at a health check-up center in Korea, and molecular identification was performed on the larvae using PCR-RFLP analysis and gene sequencing of rDNA ITS regions and mtDNA cox2. In addition, anisakid larvae (n=53) collected from the sea eel (Astroconger myriaster) were also examined for comparison with those extracted from humans. The results showed that all human samples (100%) were identified as A. pegreffii, whereas 90.7% of the samples from the sea eel were A. pegreffii with the remaining 9.3% being Hysterothylacium aduncum. Our study confirmed that A. pegreffii is the predominant species causing human anisakiasis in Korea, and this seems to be due to the predominance of this larval type in the fish (sea eels) popularly consumed by the Korean people. The possibility of human infection with H. aduncum in Korea is also suggested.

A Lung Granuloma Case Possibly Associated with a Working Environment: A Case Report

  • Seehapanya, Sankom;Chaiear, Naesinee;Ratanawatkul, Pailin;Samerpitak, Kittipan;Intarawichian, Piyapharom;Wonglakorn, Lumyai
    • Safety and Health at Work
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    • v.12 no.2
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    • pp.268-271
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    • 2021
  • Lung granulomas are uncommon in Thailand. The disease typically develops from an occupational environment and is mostly caused by infection. Herein is a case report of a female patient, aged 48, working as a nurse in an Accident and Emergency Department at a hospital. Eighteen years prior to admission the patient was diagnosed with myasthenia gravis and pulmonary tuberculosis. The chest X-ray and CT scans showed a solitary pulmonary nodule in the lower left lung. The patient received an open thoracotomy with a left lobectomy. Granulomatous and nonseptate hyphae were found in the pathology diagnosis. The patient was thus diagnosed as having a lung granuloma. The galactomannan antigen test was positive. The solitary pulmonary nodule-found from the use of a Polymerase Chain Reaction (PCR) test-was an Aspergillus spp. The fungus culture was collected from air samples. The air samples were collected by the impaction technique using a microbial air sampler. Three types of Aspergillus spp. were found as well as Penicillium spp. and Monilia sitophila. The Aspergillus spp. was a match for the patient's disease. The patient was diagnosed as having a lung granuloma possibly Aspergillus nodule which was caused by airborne Aspergillus spp. from the occupational environment.

Development of sandwich enzyme-linked immunosorbent assay for a large-scale detection of porcine transmissible gastroenteritis virus in feces

  • Oh, Yeonsu;Lee, Sang-Joon;Cho, Ho-Seong;Tark, Dongseob
    • Korean Journal of Veterinary Service
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    • v.43 no.4
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    • pp.237-244
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    • 2020
  • Porcine transmissible gastroenteritis (TGE) has been a significant cause of economic losses in pig farming industry since 1950s. Although transmissible gastroenteritis virus (TGEV) has declined in recent years, it should not be excluded because of its characteristics; the frequency of gene mutation, the mortality in piglets, and the possibility for sudden incidence. Therefore, the herd-level monitoring of the virus is important to prevent further circulation of TGE. The aim of this study is to develop a large-scale sandwich enzyme-linked immunosorbent assay (ELISA) with high specificity to rapidly detect TGEV in feces by using monoclonal antibodies (Mabs). The TGEV specific Mabs were produced in hybridoma cells. Among the Mabs belonged to the IgG class developed by this study, the final selected 8H6, 1B7, 4G3, and 1F8 were identified to have the neutralization ability against TGEV. The sandwich ELISA was established using 8H6 as a reporter antibody and 1B7 and the reported 5C8 as a capture antibody. The developed sandwich ELISA was able to distinguish TGEV from other pathogenic diarrheal agents (porcine rotavirus, porcine reovirus, porcine epidemic diarrhea virus (PEDV), E. coli, and C. perfringens) in tissue culture as well as fecal samples. And the detection rate of TGEV in feces was 80% compared with RT-PCR. The results suggested that the developed sandwich ELISA may be useful in the herd-level monitoring for effective preventive measures due to the early diagnosis of TGEV using a large amount of samples.

CircCOL1A2 Sponges MiR-1286 to Promote Cell Invasion and Migration of Gastric Cancer by Elevating Expression of USP10 to Downregulate RFC2 Ubiquitination Level

  • Li, Hang;Chai, Lixin;Ding, Zujun;He, Huabo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.938-948
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    • 2022
  • Gastric cancers (GC) are generally malignant tumors, occurring with high incidence and threatening public health around the world. Circular RNAs (circRNAs) play crucial roles in modulating various cancers, including GC. However, the functions of circRNAs and their regulatory mechanism in colorectal cancer (CRC) remain largely unknown. This study focuses on both the role of circCOL1A2 in CRC progression as well as its downstream molecular mechanism. Quantitative polymerase chain reaction (qPCR) and western blot were adopted for gene expression analysis. Functional experiments were performed to study the biological functions. Fluorescence in situ hybridization (FISH) and subcellular fraction assays were employed to detect the subcellular distribution. Luciferase reporter, RNA-binding protein immunoprecipitation (RIP), co-immunoprecipitation (Co-IP), RNA pull-down, and immunofluorescence (IF) and immunoprecipitation (IP) assays were used to explore the underlying mechanisms. Our results found circCOL1A2 to be not only upregulated in GC cells, but that it also propels the migration and invasion of GC cells. CircCOL1A2 functions as a competing endogenous RNA (ceRNA) by sequestering microRNA-1286 (miR-1286) to modulate ubiquitin-specific peptidase 10 (USP10), which in turn spurs the migration and invasion of GC cells by regulating RFC2. In sum, CircCOL1A2 sponges miR-1286 to promote cell invasion and migration of GC by elevating the expression of USP10 to downregulate the level of RFC2 ubiquitination. Our study offers a potential novel target for the early diagnosis and treatment of GC.

Development of Recombinase Polymerase Amplification Combined with Lateral Flow Strips for Rapid Detection of Cowpea Mild Mottle Virus

  • Xinyang Wu;Shuting Chen;Zixin Zhang;Yihan Zhang;Pingmei Li;Xinyi Chen;Miaomiao Liu;Qian Lu;Zhongyi Li;Zhongyan Wei;Pei Xu
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.486-493
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    • 2023
  • Cowpea mild mottle virus (CPMMV) is a global plant virus that poses a threat to the production and quality of legume crops. Early and accurate diagnosis is essential for effective managing CPMMV outbreaks. With the advancement in isothermal recombinase polymerase amplification and lateral flow strips technologies, more rapid and sensitive methods have become available for detecting this pathogen. In this study, we have developed a reverse transcription recombinase polymerase amplification combined with lateral flow strips (RT-RPA-LFS) method for the detection of CPMMV, specifically targeting the CPMMV coat protein (CP) gene. The RT-RPA-LFS assay only requires 20 min at 40℃ and demonstrates high specificity. Its detection limit was 10 copies/µl, which is approximately up to 100 times more sensitive than RT-PCR on agarose gel electrophoresis. The developed RT-RPA-LFS method offers a rapid, convenient, and sensitive approach for field detection of CPMMV, which contribute to controlling the spread of the virus.

Molecular Marker Development for the Rapid Differentiation of Black Rot Causing Xanthomonas campestris pv. campestris Race 7

  • Yeo-Hyeon Kim;Sopheap Mao;Nihar Sahu;Uzzal Somaddar;Hoy-Taek Kim;Masao Watanabe;Jong-In Park
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.494-503
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    • 2023
  • Xanthomonas campestris pv. campestris (Xcc) is a plant pathogen of Brassica crops that causes black rot disease throughout the world. At present, 11 physiological races of Xcc (races 1-11) have been reported. The conventional method of using differential cultivars for Xcc race detection is not accurate and it is laborious and time-consuming. Therefore, the development of specific molecular markers has been used as a substitute tool because it offers an accurate and reliable result, particularly a quick diagnosis of Xcc races. Previously, our laboratory has successfully developed race-specific molecular markers for Xcc races 1-6. In this study, specific molecular markers to identify Xcc race 7 have been developed. In the course of study, whole genome sequences of several Xcc races, X. campestris pv. incanae, X. campestris pv. raphani, and X. campestris pv. vesicatoria were aligned to identify variable regions like sequence-characterized amplified regions and insertions and deletions specific to race 7. Primer pairs were designed targeting these regions and validated against 22 samples. The polymerase chain reaction analysis revealed that three primer pairs specifically amplified the DNA fragment corresponding to race 7. The obtained finding clearly demonstrates the efficiency of the newly developed markers in accurately detecting Xcc race 7 among the other races. These results indicated that the newly developed marker can successfully and rapidly detect Xcc race 7 from other races. This study represents the first report on the successful development of specific molecular markers for Xcc race 7.

Diagnosing Balamuthia mandrillaris amebic meningoencephalitis in a 64-year-old woman from the Southwest of China

  • Suhua Yao;Xiaoting Chen;Lian Qian;Shizheng Sun;Chunjing Zhao;Zongkai Bai;Zhaofang Chen;Youcong Wu
    • Parasites, Hosts and Diseases
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    • v.61 no.2
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    • pp.183-193
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    • 2023
  • Balamuthia mandrillaris amebic encephalitis (BAE) can cause a fatal condition if diagnosis is delayed or effective treatment is lacking. Patients with BAE have been previously reported in 12 provinces of China, with skin lesions being the primary symptom and encephalitis developing after several years. However, a significantly lower number of cases has been reported in Southwest China. Here we report an aggressive BAE case of a 64-year-old woman farmer with a history of skin lesions on her left hand. She was admitted to our hospital due to symptoms of dizziness, headache, cough, vomiting, and gait instability. She was initially diagnosed with syphilitic meningoencephalitis and received a variety of empirical treatment that failed to improve her symptoms. Finally, she was diagnosed with BAE combined with amebic pneumonia using next-generation sequencing (NGS), qRT-PCR, sequence analysis, and imaging studies. She died approximately 3 weeks after the onset. This case highlights that the rapid development of encephalitis can be a prominent clinical manifestation of Balamuthia mandrillaris infection.

Outbreak of Hepatitis A caused by Groundwater Contamination in a Long-term Psychiatric Hospital in Korea (장기 요양 정신병원에서 지하수 오염에 의하여 발생한 A형간염 집단발병)

  • Keun-Sang Kwon;Myung Ok Lee;Hee Ju Bae;Jung Im Park;Cheon-Hyeon Kim;Ju-Hyung Lee
    • Journal of agricultural medicine and community health
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    • v.48 no.2
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    • pp.71-80
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    • 2023
  • 목적: 2017년 지하수를 식수로 사용하고 있는 한 장기요양 정신병원(H병원)에서 A형간염 환자가 집단 발병하여 이에 대한 역학조사를 실시하고 조치 결과를 기술하고자 하였다. 방법: 노출기간 동안 H병원의 근로자 및 재원 환자 234명을 대상으로 사례군 조사 디자인으로 역학조사를 실시하였고, IgM, IgG 혈청검사 및 A형간염 바이러스(HAV)에 대한 PCR검사를 시행하였다. 또한 오염원으로 의심되는 지하수, 병원에서 제공되는 식품 및 인근 저수지의 물에서 HAV 검사를 실시하였고, 검출된 HAV는 유전형 검사를 진행하였다. 결과: H병원 환자 및 직원 234명 중 IgG 양성인 168명을 제외한 66명 중 19명이 최종적으로HAV 감염자로 확인되어 감수성자 중 발병률은 28.8%로 나타났다. 환자, 지하수, 식품(석박지) 및 저수지에서 동일 유전형의 HAV가 검출되어 지하수 오염에 의한 집단발병으로 결론 내렸으나, 최초 오염원은 확인하지 못하였다. 유행 종결 선언 이후 지하수에 대한 관리로 염소소독과 UV 조사를 하였음에도 불구하고 6개월 동안 지속적으로 HAV가 검출되어 새로운 관정을 개발하여 상황을 종결하였다. 결론: 본 연구에서 지하수를 식수로 사용하는 장기요양 정신병원에서 지하수 오염에 의한 19명의 HAV 집단발병을 조사하였다. HAV 항체가 없는 대상자 중에서 HAV의 높은 발병률을 확인하였다. 지하수 수질검사에서 바이러스 검사는 포함되어 있지 않기 때문에 지하수가 HAV에 오염시 HAV 집단발병 가능성이 높고 상당기간 지속적으로 검출되기 때문에 지하수에 대한 관리지침에 바이러스 검출을 위한 방안을 추가하고 관련 법을 정비할 필요가 있다.

Alternative and Rapid Detection Methods for Wastewater Surveillance of SARS-CoV-2 (SARS-CoV-2의 하수조사를 위한 대체 및 신속 검출 방법)

  • Jesmin Akter;Bokjin Lee;Jai-Yeop Lee;Chang Hyuk Ahn;Nishimura Fumitake;ILHO KIM
    • Journal of Korean Society on Water Environment
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    • v.40 no.1
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    • pp.19-35
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    • 2024
  • The global pandemic, coronavirus disease caused by Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to the implementation of wastewater surveillance as a means to monitor the spread of SARS-CoV-2 prevalence in the community. The challenging aspect of establishing wastewater surveillance requires a well-equipped laboratory for wastewater sample analysis. According to previous studies, RT-PCR-based molecular tests are the most widely used and popular detection method worldwide. However, this approach for the detection or quantification of SARS-CoV-2 from wastewater demands a specialized laboratory, skilled personnel, expensive instruments, and a workflow that typically takes 6 to 8 hours to provide results for a few samples. Rapid and reliable alternative detection methods are needed to enable less-well-qualified practitioners to set up and provide sensitive detection of SARS-CoV-2 within wastewater at regional laboratories. In some cases, the structural and molecular characteristics of SARS-CoV-2 are unknown, and various strategies for the correct diagnosis of COVID-19 have been proposed by research laboratories. The ongoing research and development of alternative and rapid technologies, namely RT-LAMP, ELISA, Biosensors, and GeneXpert, offer a wide range of potential options not only for SARS-CoV-2 detection but also for other viruses. This study aims to discuss the effective regional rapid detection and quantification methods in community wastewater.