Osong Public Health and Research Perspectives

Original Articles

Evaluation and Comparison of Molecular and Conventional Diagnostic Tests for Detecting Tuberculosis in Korea, 2013: Sang-Hee Park, Chang-Ki Kim, Hye-Ran Jeong, Hyunjin Son, Seong-Han Kim, Mi-Sun Park; Osong Public Health Res Perspect. 2014;5(Suppl):S3-S7. Published online December 31, 2014; DOI: https://doi.org/10.1016/j.phrp.2014.10.006

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Objectives
A fast and accurate diagnosis is necessary to control and eliminate tuberculosis (TB). In Korea, TB continues to be a serious public health problem. In this study, diagnostic tests on clinical samples from patients suspected to have TB were performed and the sensitivity and specificity of the various techniques were compared. The main objective of the study was to compare various diagnostic tests and evaluate their sensitivity and specificity for detecting tuberculosis.
Methods
From January 2013 to December 2013, 170,240 clinical samples from patients suspected to have TB were tested with smear microscopy, acid-fast bacilli culture, and real-time polymerase chain reaction (PCR). The test results were compared and data were analyzed.
Results
A total of 8216 cultures tested positive for TB (positive detection rate, 4.8%). The contamination rate in the culture was 0.6% and the isolation rate of nontuberculous mycobacteria was 1.0%. The sensitivity and specificity of smear microscopy were 56.8% and 99.6%, respectively. The concordance rate between the solid and liquid cultures was 92.8%. Mycobacterium isolates were not detected in 0.4% of the cases in the liquid culture, whereas no Mycobacterium isolates were detected in 6.8% of the cases in the solid culture. The sensitivity and specificity of real-time PCR for the solid culture were 97.2% and 72.4%, respectively, whereas the corresponding data for the liquid culture were 93.5% and 97.2%.
Conclusion
The study results can be used to improve existing TB diagnosis procedure as well as for comparing the effectiveness of the assay tests used for detecting Mycobacterium tuberculosis isolates.

Citations

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The Xpert® MTB/RIF diagnostic test for pulmonary and extrapulmonary tuberculosis in immunocompetent and immunocompromised patients: Benefits and experiences over 2 years in different clinical contexts
Ana Paula de Oliveira Tomaz, Sonia Mara Raboni, Gislene Maria Botão Kussen, Keite da Silva Nogueira, Clea Elisa Lopes Ribeiro, Libera Maria Dalla Costa, Padmapriya P. Banada
PLOS ONE.2021; 16(3): e0247185. CrossRef
Comparative performance of the laboratory assays used by a Diagnostic Laboratory Hub for opportunistic infections in people living with HIV
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Multiplex PCR is a Rapid, Simple and Cheap Method for Direct Diagnosis of M. tuberculosis from Sputum Samples
Tarig M.S. Alnour, Faisel Abuduhier , Mohammed Khatatneh , Fahad Albalawi , Khalid Alfifi , Bernard Silvala
Infectious Disorders - Drug Targets .2020; 20(4): 495. CrossRef
Naked eye detection of the Mycobacterium tuberculosis complex by recombinase polymerase amplification—SYBR green I assays
Nuntita Singpanomchai, Yukihiro Akeda, Kazunori Tomono, Aki Tamaru, Pitak Santanirand, Panan Ratthawongjirakul
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Retropharyngeal SOL: An unusual presentation of a multifaceted entity
Poojan Agarwal, Manju Kaushal, Shruti Dogra, Ankur Gupta, Nishi Sharma
CytoJournal.2018; 15: 12. CrossRef
Rapid Detection of Rifampicin- and Isoniazid-Resistant Mycobacterium tuberculosis using TaqMan Allelic Discrimination
Davood Darban-Sarokhalil, Mohammad J. Nasiri, Abbas A.I. Fooladi, Parvin Heidarieh, Mohammad M. Feizabadi
Osong Public Health and Research Perspectives.2016; 7(2): 127. CrossRef
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Lon Mutant of Brucella abortus Induces Tumor Necrosis Factor-Alpha in Murine J774.A1 Macrophage: Sungdo Park, Young-Sill Choi, Sang-Hee Park, Young-Rok Kim, Hyuk Chu, Kyu-Jam Hwang, Mi-Yeoun Park; Osong Public Health Res Perspect. 2013;4(6):301-307. Published online December 31, 2013; DOI: https://doi.org/10.1016/j.phrp.2013.10.002

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Abstract PDF

Objectives
The objective of this study was to isolate a Brucella lon mutant and to analyze the cytokine response of B. lon mutant during macrophage infection.
Methods
A wild-type Brucella abortus strain was mutagenized by Tn5 transposition. From the mouse macrophage J774.A1 cells, total RNA was isolated at 0 hours, 6 hours, 12 hours, and 24 hours after infection with Brucella. Using mouse cytokine microarrays, we measured transcriptional levels of the cytokine response, and validated our results with a reverse transcriptase-polymerase chain reaction (RT-PCR) assay to confirm the induction of cytokine messenger RNA (mRNA).
Results
In host J774.A1 macrophages, mRNA levels of T helper 1 (Th1)-type cytokines, including tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin-2 (IL-2), and IL-3, were significantly higher in the lon mutant compared to wild-type Brucella and the negative control. TNF-α levels in cell culture media were induced as high as 2 μg/mL after infection with the lon mutant, a greater than sixfold change.
Conclusion
In order to understand the role of the lon protein in virulence, we identified and characterized a novel B. lon mutant. We compared the immune response it generates to the wild-type Brucella response in a mouse macrophage cell line. We demonstrated that the B. lon mutants induce TNF-α expression from the host J774.A1 macrophage.

Citations

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Structure, Substrate Specificity and Role of Lon Protease in Bacterial Pathogenesis and Survival
Perumalraja Kirthika, Khristine Kaith Sison Lloren, Vijayakumar Jawalagatti, John Hwa Lee
International Journal of Molecular Sciences.2023; 24(4): 3422. CrossRef
ClpP protease modulates bacterial growth, stress response, and bacterial virulence in Brucella abortus
Dongjie Sun, Yufu Liu, Xiaowei Peng, Hao Dong, Hui Jiang, Xuezheng Fan, Yu Feng, Jiali Sun, Kun Han, Qiang Gao, Jianrui Niu, Jiabo Ding
Veterinary Research.2023;[Epub] CrossRef
Brucella abortus Encodes an Active Rhomboid Protease: Proteome Response after Rhomboid Gene Deletion
María Inés Marchesini, Ansgar Poetsch, Leticia Soledad Guidolín, Diego J. Comerci
Microorganisms.2022; 10(1): 114. CrossRef
Proteomics of Brucella
Ansgar Poetsch, María Inés Marchesini
Proteomes.2020; 8(2): 8. CrossRef
Outer Membrane Vesicles From Brucella melitensis Modulate Immune Response and Induce Cytoskeleton Rearrangement in Peripheral Blood Mononuclear Cells
Eric Daniel Avila-Calderón, Olín Medina-Chávez, Leopoldo Flores-Romo, José Manuel Hernández-Hernández, Luis Donis-Maturano, Ahidé López-Merino, Beatriz Arellano-Reynoso, Ma. Guadalupe Aguilera-Arreola, Enrico A. Ruiz, Zulema Gomez-Lunar, Sharon Witonsky,
Frontiers in Microbiology.2020;[Epub] CrossRef
RNA-seq reveals the critical role of Lon protease in stress response and Brucella virulence
Yufu Liu, Hao Dong, Xiaowei Peng, Qiang Gao, Hui Jiang, Guanlong Xu, Yuming Qin, Jianrui Niu, Shijing Sun, Peng Li, Jiabo Ding, Ruiai Chen
Microbial Pathogenesis.2019; 130: 112. CrossRef
Brucella Downregulates Tumor Necrosis Factor-α to Promote Intracellular Survival via Omp25 Regulation of Different MicroRNAs in Porcine and Murine Macrophages
Xiaomao Luo, Xiujuan Zhang, Xingchen Wu, Xuefeng Yang, Cong Han, Zhengyu Wang, Qian Du, Xiaomin Zhao, Shan-Lu Liu, Dewen Tong, Yong Huang
Frontiers in Immunology.2018;[Epub] CrossRef

Evaluation and Selection of Multilocus Variable-Number Tandem-Repeat Analysis Primers for Genotyping Brucella abortus Biovar 1 Isolated from Human Patients: Subok Lee, Kyu-Jam Hwang, Mi-Yeoun Park, Seon-Do Hwang, Hee-Youl Chai, Hyuk Chu, Sang-Hee Park; Osong Public Health Res Perspect. 2013;4(5):265-270. Published online October 31, 2013; DOI: https://doi.org/10.1016/j.phrp.2013.09.005

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Abstract PDF

Objectives
Brucellosis is the most common bacterial zoonosis in the world. Multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) is a molecular method for genotyping bacterial species. Brucella abortus biovar I was isolated from most of the brucellosis-suspected patients in Korea. This study was conducted to investigate the ability of various MLVA primers that are used for molecular typing B. abortus isolates and for analyzing their epidemiological data.
Methods
A total of 80 human isolates of B. abortus biovar I isolated from human patients and the reference strain were used for MLVA. Genetic diversity was determined by calculating the Simpson's diversity index (DI) of each VNTR locus. The Brucella strains were subcultured 30 times to determine the stability of each locus. The DNA of the strains cultivated in each passage was extracted and subjected to MLVA for further investigation.
Results
The 15 VNTR loci were selected based on high DI values. The DIs of the 15 VNTR loci showed considerable discrimination power ranging from 59% for Bruce 43 to 87% for Bruce 22. Bruce 09, Bruce 11, Bruce 16, Bruce 42, and Bruce 43 were confirmed to remain stable in vitro among the 15 VNTR loci selected.
Conclusion
The results of this study suggest that the five loci subsets may be a useful epidemiological tool for investigating B. abortus biovar 1 outbreak.

Citations

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Brucella abortus: Current Research and Future Trends
Tariq Jamil, Falk Melzer, John Njeru, Hosny El-Adawy, Heinrich Neubauer, Gamal Wareth
Current Clinical Microbiology Reports.2017; 4(1): 1. CrossRef

Article

Application of the Microagglutination Test for Serologic Diagnosis of Human Brucellosis: Sang-Hee Park, Yoo-Hoon Lee, Hyuk Chu, Seon-Do Hwang, Kyu-Jam Hwang, Hee-Yeol Choi, Mi-Yeoun Park; Osong Public Health Res Perspect. 2012;3(1):19-23. Published online December 31, 2011; DOI: https://doi.org/10.1016/j.phrp.2012.01.003

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Abstract PDF

Objectives
Brucellosis is one of the most common zoonoses in the world, and occurs mainly in farmers, slaughterhouse workers, and veterinarians via direct or indirect contact with infected animals or their products. The clinical symptoms of human brucellosis are nonspecific, such as fever, headache, chills, and sweating. Diagnosis and treatment of brucellosis requires laboratory tests. Although the serum tube agglutination test (SAT) is the standardized gold method, it is laborious, time consuming, and requires a number of reagents. A microagglutination test (MAT) variant of the SAT or enzyme-linked immunosorbent assay (ELISA) is recommended for serological diagnoses. For the simple and rapid diagnosis of brucellosis, the MAT was standardized using samples for the SAT to define positive and negative categories, and we then compared the sensitivity and specificity of the MAT and ELISA.
Methods
Thirty SAT-positive sera and 60 SAT-negative sera were used in this study. Antibody titers of ≥1:160 were considered positive readings in both the SAT and MAT. Brucella abortus antigens and Brucella-positive control antiserum were used in the SAT and MAT. ELISAs of IgM and IgG were performed according to the manufacturers’ instructions.
Results
The titers of the MAT differed according to antigen concentration. The optimal concentration of B abortus antigen was determined to compare the sensitivity and specificity between the MAT and SAT. The sensitivity and specificity of the MAT were 93.3% and 96.7%, respectively, for IgG with reference to ELISA, and 96.7% and 98.3%, respectively, for IgM.
Conclusions
The optimal concentration of antigen for the MAT was 1:10. The MAT is less time consuming and requires less antigen and serum than the SAT. The results of the MAT showed good agreement with those of ELISA. The results of this study suggest that the MAT could be useful for diagnosis of brucellosis.

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Original Article

Serological Detection of Lyme Borreliosis Agents in Patients From Korea, 2005–2009: Sang-Hee Park, Kyu-Jam Hwang, Hyuk Chu, Mi-Yeoun Park; Osong Public Health Res Perspect. 2011;2(1):29-33. Published online June 30, 2011; DOI: https://doi.org/10.1016/j.phrp.2011.04.004

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Abstract PDF

Objectives
Laboratory tests are now being used to identify seropositive cases in patients suspected of having a Lyme borreliosis (LB) infection. From 2005 to 2009, we analyzed the serological and epidemiological characteristics of 53 LB positive cases in Korea using immunoblot assay.
Methods
During the five-year study period, a total of 1897 serum samples from suspected LB cases were referred to us for further laboratory diagnosis. The bacterial strains Borrelia afzeli pKo, Borrelia garinii 935T and Borrelia burgdorferi B31 were used for indirect immunofluorescent antibody assay. Immunoblot assay was performed using the recomBlot Borrelia.
Results
Based on the information from the clinicians, the main symptoms of LB infection were rash and fever (66.0%), neurological symptoms (30.2%), and arthritis (5.7%). Of the 53 cases, 16 (30.2%) were infected abroad and the remaining 37 cases (69.8%) were suspected to have been infected in Korea. Immunoblot assays detected high levels of the antigens p41 (FlaB) of B. burgdorferi and OspC of B. garinii in infected samples.
Conclusions
The causative bacteria of LB were not isolated from humans yet but from vector ticks and rodents in Korea, and a few cases were reported with serological diagnosis. Our results suggest that LB is present in all areas of Korea and indicate that B. garinii and B. burgdorferi may be the predominant bacteria in patients with LB. However, further studies are needed to isolate and identify the causative bacteria for LB in patients.

Citations

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Seung-Ho Lee, Sung-Tae Chong, Heung-Chul Kim, Terry A Klein, Kyungmin Park, Jingyeong Lee, Jeong-Ah Kim, Won-Keun Kim, Jin-Won Song, Sarah Hamer
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Seung-Hun Lee, Youn-Kyoung Goo, Paul John L. Geraldino, Oh-Deog Kwon, Dongmi Kwak
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