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Development and Evaluation of a Rapid Dipstick Assay for Serodiagnosis of Bovine Brucellosis
Hannah Leah Tadeja Simborio,Alisha Wehdnesday Bernardo Reyes,Huynh Tan Hop,Lauren Togonon Arayan,Kwang Myun Cheong,김석 경상대학교 농업생명과학연구원 2016 농업생명과학연구 Vol.50 No.6
Fast, cheap and sufficient serodiagnostic tools needs to be developed for the early detectionof brucellosis. Currently the tools cannot differentiate an active infection from vaccinated, norcan it differentiate other bacterial infections with lipopolysaccharides, especially Yersiniainfections. In this study, we purified recombinant outer membrane protein 10 and 28(rOmp10,rOmp28), and a dipstick assay(indirect or sandwich) was constructed with single(rOmp10 orrOmp28) and combined rOmps(rOmp10 and rOmp28) from Brucella(B.) abortus 544 to evaluatebovine Brucella positive serum collected during the beginning of the Korean outbreak from2006 to 2015. In application with single rOmp, rOmp10(70%; indirect, 92.11%; sandwichdipstick) and rOmp28(72.5%; indirect, 86.84%; sandwich dipstick) had comparable results. Inaddition, results indicated that dipstick with combined rOmps(rOmps10 and rOmp28) weresuperior in detecting positive serum samples, at 85% indirect and 100% sandwich dipstick. Surprisingly, the results were the same in detecting negative results at 97.78% for both singleand combined indirect dipsticks. The dipstick tools with rOmp10 and rOmp28 would be usefulfor a rapid screen method for bovine brucellosis.
( Hannah Leah Tadeja Simborio ),( Alisha Wehdnesday Bernardo Reyes ),( Huynh Tan Hop ),( Lauren Togonon Arayan ),( Wongi Min ),( Hu Jang Lee ),( Jin Ju Lee ),( Hong Hee Chang ),( Suk Kim ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.3
Brucellosis affects a wide range of host species, including humans and many livestock animals. Chronic infections of the disease make antibiotic treatment costly, and the current vaccine used in livestock has not been approved for human use. This study investigated the possible use of the Brucella abortus outer membrane protein A (OmpA) as a candidate subunit vaccine in an infected mouse model. The ompA gene was cloned and overexpressed, and the recombinant OmpA (rOmpA) protein fused to maltose binding protein (MBP) was purified in Escherichia coli. Immunogenicity was verified through western blotting, and mice were immunized and challenged to evaluate its protective effect. Mice treated with rOmpA exhibited induced humoral and host cell-mediated responses, with a significant increase in immunoglobulin G (IgG1 and IgG2a) and cytokine levels, especially TNF-α and IL-12, compared with the control groups treated with either MBP or PBS. In conclusion, rOmpA should be highly considered as a future subunit vaccine for brucellosis, and further studies regarding rOmpA and its protective ability are suggested.
Review Article : Strategies for the development of an effective vaccine against Brucellosis
( Hannah Leah Tadeja Simborio ),( Alisha Wehdnesday Bemardo Reyes ),( Huynh Tan Hop ),( Lauren Togonon Arayan ),( Won Gi Min ),( Hu Jang Lee ),( Hong Hee Chang ),( Suk Kim ) 한국예방수의학회 2014 예방수의학회지 Vol.38 No.2
Brucellosis is a notorious zoonotic disease with global implications. Efforts to control the spread of the disease have been restricted to the agricultural livestock. Increasing incidences of accidental human infection have motivated researches to start working on alternative vaccines. At present, live attenuated vaccines are the only accepted type of vaccines used in developed countries for the prevention of brucellosis. Although serodiagnosis is occasionally unreliable, some countries have already claimed to have eradicated the disease, based on this testing. Live attenuated vaccines are not suitable for use in pregnant and immune-depressed animals. Moreover, these vaccines are not tolerated in humans. Therefore, many researches have been striving to discover alternative methods of vaccination. Most research has focused on the generation of subcellular, subunit, and DNA vaccines that are as efficient as the live attenuated vaccines. At present, none of the available vaccines has been able to replace the live attenuated vaccines. Therefore, additional research is necessary in order to discover a new brucellosis vaccine that is suitable for human use.
Lee, Jin Ju,Simborio, Hannah Leah,Reyes, Alisha Wehdnesday Bernardo,Kim, Dae Geun,Hop, Huynh Tan,Min, Wongi,Her, Moon,Jung, Suk Chan,Yoo, Han Sang,Kim, Suk BioMed Central 2015 VETERINARY RESEARCH Vol.46 No.-
<P>Brucellosis is a vital zoonotic disease caused by <I>Brucella</I>, which infects a wide range of animals and humans. Accurate diagnosis and reliable vaccination can control brucellosis in domestic animals. This study examined novel immunogenic proteins that can be used to detect <I>Brucella abortus</I> infection or as an effective subcellular vaccine. In an immunoproteomic assay, 55 immunodominant proteins from <I>B. abortus</I> 544 were observed using two dimensional electrophoresis (2DE) and immunoblot profiles with antisera from <I>B. abortus</I>-infected cattle at the early (week 3), middle (week 7), and late (week 10) periods, after excluding protein spots reacting with antisera from <I>Yersinia enterocolitica</I> O:9-infected and non-infected cattle. Twenty-three selected immunodominant proteins whose spots were observed at all three infection periods were identified using MALDI-MS/MS. Most of these proteins identified by immunoblot and mass spectrometry were determined by their subcellular localization and predicted function. We suggest that the detection of prominent immunogenic proteins during the infection period can support the development of advanced diagnostic methods with high specificity and accuracy; subsidiarily, these proteins can provide supporting data to aid in developing novel vaccine candidates.</P>
Lee, Jin Ju,Simborio, Hannah Leah,Reyes, Alisha Wehdnesday Bernardo,Kim, Dae Geun,Hop, Huynh Tan,Min, Wongi,Her, Moon,Jung, Suk Chan,Yoo, Han Sang,Kim, Suk Published by Elsevier/North Holland on behalf of t 2014 FEMS microbiology letters Vol.357 No.2
<P>Brucellosis is a major zoonotic disease caused by pathogens of the genus Brucella. The eradication of brucellosis in domestic animals, associated with the prevention of human infection, can be attained through accurate diagnosis. However, the conventional serological diagnosis of brucellosis has limitations, particularly in detecting the infection period. Accordingly, the aim of this study was to determine reliable immunogenic proteins to detect Brucella abortus infection according to time course responses to aid in the appropriate management of this disease. Proteomic identification through two-dimensional electrophoresis (2DE), followed by immunoblotting, revealed 13, 24, and 55 immunodominant B.?abortus 544 proteins that were reactive to sera from experimentally infected mice at early (10?days), middle (30?days), and late (60?days) infection periods, respectively. After excluding several spots reactive to sera from Yersinia enterocolitica O:9-infected and noninfected mice, 17 of the 67 immunodominant proteins were identified through MALDI-TOF MS. Consequently, the identified proteins showed time course-dependent immunogenicity against Brucella infection. Thus, the results of this study suggest that the production of immunogenic proteins during infection periods improves the diagnosis and discovery of vaccine candidates.</P>
김대근,이진주,Hannah Leah Simborio,Alisha Wehdnesday Bernardo Reyes,민원기,이후장,장홍희,김석 경상대학교 농업생명과학연구원 2013 농업생명과학연구 Vol.47 No.2
Brucellosis is an important bacterial zoonosis in humans and domestic animals. Brucella spp. are taken up, and survive within non-professional and professional phagocytes. In common belief, diabetes mellitus increases susceptibility to pathogenic infection. In this study, Brucella (B.) abortus was inoculated into a diabetic animal model, db/db mice, in order to show the course of brucellosis in diabetic state. The liver proliferation, bacterial burden of the liver, level of cytokines in serum and macrophage migration into liver, were investigated at 14 days post-infection. In comparison with the uninfected control mice, the results revealed that the weight of the liver of infected db/db mice was higher but with lower bacterial load in this organ. The level of MCP-1 mRNA expression in the liver was lower, the levels of IL-12p70, IL-10, TNF-α and IFN-γ in serum was significantly higher and the macrophages migration was significantly lower in infected mice than in the control group. In conclusion, this present study suggested that MCP-1 suppression by B. abortus infection may inhibit the macrophages migration, and consequently may induce to abrogate the bacterial survival in db/db mouse liver. Furthermore, the increased inflammatory cytokines may contribute to inhibition of B. abortus proliferation in diabetic mice.
Antibacterial effects of Coptis chinensis Franch against Brucella abortus
김대근,Hannah Leah Tadeja Simborio,Alisha Wehdnesday Bernardo Reyes,민원기,이후장,이진주,장홍희,김동혁,김석 경상대학교 농업생명과학연구원 2014 농업생명과학연구 Vol.48 No.1
Brucellosis is an important bacterial zoonotic infection worldwide responsible for economic losses in livestock industries which represents a considerable and increasing public health burden. The disease is caused by Brucella which possess the ability to invade and replicate within phagocytes and may result to chronic infection, leading to difficulty in medical therapy of the disease. The treatment for brucellosis employs conventional principles that have been applied for a long time and a combination of antibiotics is currently used due to low efficacy of monotherapies. However, vital effects with respect to health and safety are neglected. Nevertheless, the preventive methods and treatments for brucellosis using traditional medicine have not yet been thoroughly studied, hence, alternative therapies such as the use of natural plant extracts as traditional medicine that are safe, efficient and economical should be explored in order to identify candidates that eliminate complications due to brucellosis. Coptis chinensis Franch (Huanglian) is a traditional Chinese medicinal herb and its extract has been known to possess strong antibacterial activity. In this study, the antibacterial effects of C. chinensis Franch extracts, and the major components of the herb namely berberine and palmatine, were investigated on B. abortus. The C. chinensis Franch ethanol extracts (CCFE) showed bactericidal effects at 1,000 μg/ml concentration and berberine at 100 μg/ml concentration. However, C. chinensis Franch and its components did not affect invasion and intracellular growth of B. abortus in RAW 264.7 cells. In conclusion, C. chinensis Franch ethanol extracts, water extract and its major components such as berberine and palmatine would be a beneficial antimicrobial agent without affecting phagocytic pathway within macrophages, and further study for the precise mechanisms of antibacterial effect would be necessary.