• Title/Summary/Keyword: detection kit

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Infection and Rapid Detection of Perkinsus sp. In Cultured Babyneck Clam, Ruditapes philippinarum from Western Coast of Korea (서해안 양식 바지락에 발생한 Perkinsus sp. 감염증과 신속검출)

  • Choi, Dong-Lim;Kwon, Jung-No;Park, Sung-Woo
    • Journal of fish pathology
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    • v.11 no.1
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    • pp.69-76
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    • 1998
  • An apicomplexan parasite, Perkinsus sp. was observed from the cultured baby clams, Ruditapes philippinarum, collected from the coast of Kochang and Taean (South Korea), where it caused seasonal mortality of clams. Several milky-white cysts were observed on the surface of gill and visceral mass of parasitised clams. The trophozoites of parasite had eccentric nucleus and proliferated by schizogony in gill, mantle, hepatopancrease and reproductive tissues, resulting in the formation of granuloma and the intensive infiltration of hemocytes in the tissues. During incubation in FTM, trophozoites increased in size, resulting in prezoosporangia which appeared as round black spheres when colored with Lugols iodine solution. The prevalence of Perkinsus sp. in clams was Kochang, 73.1%; Taean, 94.8% (during 9-mo. survey) and showed size-dependent infection. Hemacolor kit was useful to reduce time for diagnosis of the trophozoite of Perkinsus sp. that has been responsible of massive motalities in the clam.

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A Novel Anti-Microbial Peptide from Pseudomonas, REDLK Induced Growth Inhibition of Leishmania tarentolae Promastigote in Vitro

  • Yu, Yanhui;Zhao, Panpan;Cao, Lili;Gong, Pengtao;Yuan, Shuxian;Yao, Xinhua;Guo, Yanbing;Dong, Hang;Jiang, Weina
    • Parasites, Hosts and Diseases
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    • v.58 no.2
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    • pp.173-179
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    • 2020
  • Leishmaniasis is a prevalent cause of death and animal morbidity in underdeveloped countries of endemic area. However, there is few vaccine and effective drugs. Antimicrobial peptides are involved in the innate immune response in many organisms and are being developed as novel drugs against parasitic infections. In the present study, we synthesized a 5-amino acid peptide REDLK, which mutated the C-terminus of Pseudomonas exotoxin, to identify its effect on the Leishmania tarentolae. Promastigotes were incubated with different concentration of REDLK peptide, and the viability of parasite was assessed using MTT and Trypan blue dye. Morphologic damage of Leishmania was analyzed by light and electron microscopy. Cellular apoptosis was observed using the annexin V-FITC/PI apoptosis detection kit, mitochondrial membrane potential assay kit and flow cytometry. Our results showed that Leishmania tarentolae was susceptible to REDLK in a dose-dependent manner, disrupt the surface membrane integrity and caused parasite apoptosis. In our study, we demonstrated the leishmanicidal activity of an antimicrobial peptide REDLK from Pseudomonas aeruginosa against Leishmania tarentolae in vitro and present a foundation for further research of anti-leishmanial drugs.

Design of a Smart Safety Vest Incorporated With Metal Detector Kits for Enhanced Personal Protection

  • Rajendran, Salini D.;Wahab, Siti N.;Yeap, Swee P.
    • Safety and Health at Work
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    • v.11 no.4
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    • pp.537-542
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    • 2020
  • Background: Personal protective equipment (PPE) has been designed in such a way to reduce accident rates. Unfortunately, existing PPE is rather ineffective as it is not able to provide warning signals when hazard is around. The integration of intelligent systems is envisaged to increase the efficiency of existing PPE. Methods: This project designed a safety vest incorporated with metal detectors which can provide immediate warning to the field workers when there is metal hazard around. This product has greater freedom of design via smart manufacturing as it involves the assembly of few commercially available parts into a single entity. Briefly, the metal detector is a do it yourself (DIY) kit, and the safety vest is purchasable from any local market. The DIY kit was connected to a copper coil and being sewed into the safety vest. Results: The metal detector induces beeping sound when there is metal hazard around. A total of 121 engineering students were introduced to the prototype before being requested to answer a survey associated with the design. Respondents have rated >3.00/5.00 for the design simplicity, ease of usage, and light weight. Meanwhile, respondents suggested that the design should be further improved by increasing the metal detection range. Conclusion: It is envisaged that the introduction of this smart safety vest will allow the workers to carry out their duties securely by reducing the accident rates. Particularly, such design is expected to reduce workplace accident especially during night time at construction sites where the visibility is low.

Microbiological Quality of Raw and Cooked Foods in Middle and High School Food Service Establishments (서울시 일부 중.고등학교의 급식용 식재료 및 조리식품의 미생물학적 품질)

  • Kim, Myung-Hee;Shin, Weon-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.10
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    • pp.1343-1356
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    • 2008
  • The evaluation of microbiological quality for school food samples collected from 19 selected middle and high schools located in Seoul was undertaken. Eighty-nine food samples consisting of 38 non-pretreated vegetables, 13 pre-washed and cut vegetables, 9 meats and poultry, 3 fish and shellfish, 7 dried fish, and shellfish and 20 processed foods were collected. Aerobic plate count, total coliforms, and Escherichia coli (E. coli ) were detected using $Petrifilm^{TM}$, and the food-borne pathogens were screened by multiplex PCR with species-specific primer sets. Sequentially, the quantitative and confirmative test of the food-borne pathogens were carried out with the selective media and biochemical kits. The contamination of coliform counts was observed on the pre-washed vegetables ($3.4{\sim}4.3\;log\;CFU/g$) and meats ($2.2{\sim}4.3\;log\;CFU/g$). Also, the cooked foods were heavily contaminated with coliform, ranging from 1.0 to $5.5\;log\;CFU/g$. E. coli counts were found in 16 raw and cooked food samples, exceeding the microbiological standards for the guideline of safety management for school foods. Through PCR detection, B acillus cereus was detected in 32 raw and cooked foods, and quantitatively found in pre-washed carrot, radish, and pan-broiled dried shrimp and filefish ranging from $2.3{\sim}3.6\;log\;CFU/g$, respectively. E. coli O157:H7 was detected on frozen pork sample and was confirmed with API kit. Campylobacter jejuni was found in 3 ready-to-eat type vegetables. Vibrio parahaemolyticus were found in 4 pre-washed vegetables and 2 cooked foods, indicating unsatisfactory quality based upon the microbiological standards of ready-to-eat vegetables and cooked foods by Korea Food and Drug Administration. Salmonella spp. was detected in frozen chicken sample and confirmed by API kit and latex antisera agglutination.

Comparative Assessment of Diagnostic Performances of Two Commercial Rapid Diagnostic Test Kits for Detection of Plasmodium spp. in Ugandan Patients with Malaria

  • Bahk, Young Yil;Park, Seo Hye;Lee, Woojoo;Jin, Kyoung;Ahn, Seong Kyu;Na, Byoung-Kuk;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
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    • v.56 no.5
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    • pp.447-452
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    • 2018
  • Prompt diagnosis of malaria cases with rapid diagnostic tests (RDTs) has been widely adopted as an effective malaria diagnostic tool in many malaria endemic countries, primarily due to their easy operation, fast result output, and straightforward interpretation. However, there has been controversy about the diagnostic accuracy of RDTs. This study was conducted to evaluate the diagnostic performances of the 2 commercially available malaria RDT kits, RapiGEN Malaria Ag Pf/Pv (pLDH/pLDH) and Asan $EasyTest^{TM}$ Malaria Ag Pf/Pv (HRP-2/pLDH) for their abilities to detect Plasmodium species in blood samples collected from Ugandan patients with malaria. To evaluate the diagnostic performances of these 2 RDT kits, 229 blood samples were tested for malaria infection by microscopic examination and a species-specific nested polymerase chain reaction. The detection sensitivities for P. falciparum of Malaria Ag Pf/Pv (pLDH/pLDH) and Asan $EasyTest^{TM}$ Malaria Ag Pf/Pv (HRP-2/pLDH) were 87.83% and 89.57%, respectively. The specificities of the 2 RDTs were 100% for P. falciparum and mixed P. falciparum/P. vivax infections. These results suggest that the 2 RDT kits showed reasonable levels of diagnostic performances for detection of the malaria parasites from Ugandan patients. However, neither kit could effectively detect P. falciparum infections with low parasitaemia (<$500parasites/{\mu}l$).

Development of a One-Step PCR Assay with Nine Primer Pairs for the Detection of Five Diarrheagenic Escherichia coli Types

  • Oh, Kyung-Hwan;Kim, Soo-Bok;Park, Mi-Sun;Cho, Seung-Hak
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.862-868
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    • 2014
  • Certain Escherichia coli (E. coli) strains have the ability to cause diarrheal disease. Five types of diarrheagenic E. coli have been identified, including EHEC, ETEC, EPEC, EAEC, and EIEC. To detect these five diarrheagenic types rapidly, we developed a one-step multiplex PCR (MP-PCR) assay using nine primer pairs to amplify nine virulence genes specific to the different virotypes, with each group being represented (i.e., stx1 and stx2 for EHEC, lt, sth, and stp for ETEC, eaeA and bfpA for EPEC, aggR for EAEC, and ipaH for EIEC). The PCR primers were constructed using MultAlin. The sensitivity and specificity of the constructed multiplex PCR primers were measured using DNA isolated from diarrheagenic E. coli strains representing each group. The limits of detection were as follows: $5{\times}10^1CFU/ml$ for EHEC, $5{\times}10^3CFU/ml$ for ETEC expressing lt and sth, $5{\times}10^4CFU/ml$ for ETEC expressing stp, $5{\times}10^2CFU/ml$ for EPEC, $5{\times}10^4CFU/ml$ for EAEC, and $5{\times}10^2CFU/ml$ for EIEC. To confirm the specificity, C. jejuni, C. perfringens, S. Typhimurium, V. parahaemolyticus, L. monocytogenes, Y. enterocolitica, B. cereus, and S. aureus were used as negative controls, and no amplification was obtained for these. Moreover, this kit was validated using 100 fecal samples from patients with diarrhea and 150 diarrheagenic E. coli strains isolated in Korea. In conclusion, the multiplex PCR assay developed in this study is very useful for the rapid and specific detection of five diarrheagenic E. coli types. This single-step assay will be useful as a rapid and economical method, as it reduces the cost and time required for the identification of diarrheagenic E. coli.

Comparative analysis of serological tests and fecal detection in the diagnosis of Mycobacterium avium subspecies paratuberculosis infection

  • Park, Hong-Tae;Ha, Seungmin;Park, Hyun-Eui;Shim, Soojin;Hur, Tai Young;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.60 no.3
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    • pp.117-122
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    • 2020
  • Johne's disease (JD) caused by Mycobacterium avium subspecies paratuberculosis (MAP) is a chronic, wasting infectious disease in ruminants that causes enormous economic losses to the dairy and beef cattle industries. The most effective way to eradicate JD is to detect infected individuals as early as possible and remove them from the herd. However, it is difficult to detect infected individuals early with the currently using diagnostic methods. Two serological diagnostic kits commercially used worldwide and a fecal detection test were compared using 298 serum samples and feces of cattle in this study to present an efficient diagnostic method. Although there was a high correlation between the 2 serological diagnostic kits (R2 = 0.7473), kit A showed a higher serological positive rate. However, the correlation between fecal tests and serological diagnosis was very low. MAP was also detected in fecal tests in many serologically negative individuals. In the periodical diagnosis of JD, MAP was detected in the feces of only cows with the higher antibody titer to MAP. These results suggest that for effective eradication of JD, early detection of infected individuals by fecal tests together with the serological tests currently in use and by removal of infected individuals are needed.

Telomerase Activity in Benign and Malignant Thyroid Diseases (갑상선 결절의 Telomerase 활성도에 대한 분석)

  • Park Cheong-Soo;Chung Woong-Youn;Lee Mi-Kyung;Chang Hang-Suk
    • Korean Journal of Head & Neck Oncology
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    • v.14 no.2
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    • pp.199-205
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    • 1998
  • Objective: Telomerase, a specialized ribonucleoprotein polymerase associated with cellular immortality, is expressed by most malignant cells and is inactive in most normal somatic cells. The assays of telomerase activity in various tumors have provided both diagnostic and prognostic information. This study was carried out to determine whether telomerase activity could be useful in distinguishing benign and malignant thyroid diseasees. Materials & Methods: Telomerase activity was determined using Oncor $TRAP_{EZE}^{TM}ELISA$ Telomerase Detection Kit for performing PCR-based telomeric repeat amplification protocol (TRAP) assay followed by ELISA detection in both normal and tumor tissues of 23 adenomatous hyperplasias, 12 follicular adenomas, 4 follicular carcinomas, 16 papillary carcinomas, 4 Hashimoto's thyroiditises and 3 malignant lymphomas. We also examined all cases microscopically to review the status of lymphoid infiltrate. Results: Of the 62 cases, extensive lymphoid infiltrates were contained in 20 tumor tissues(4 Hashimoto's thyroiditises, 3 malignant lymphomas, 6 adenomatous hyperplasias and 7 papillary carcinomas), all of which showed positive telomerase activity. All the normal tissues without lymphoid infiltrates(n=43) did not express telomerase activity. Of 42 tumor tissues without lymphoid infiltrates, 37(88.0%) showed positive telomerase activity: 13 of 17 adenomatous hyperplasias(76.5%), 11 of 12 follicular adenomas(91.7%), 4 of 4 follicular carcinomas(100.0%) and 9 of 9 papillary carcinomas(100.0%). Conclusions: Our methods showed high sensitivity in the detection of telomerase activity and the exclusion of lymphoid infiltrates may be important in telomerase assay. In our work, the measurement of telomerase activity was not useful in distinguishing benign and malignant thyroid diseases.

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Detection of Helicobacter pylori Infection Rate in Resected Gastric Cancer Patients (위절제술을 받은 위암 환자의 Helicobacter pylori 감염률 측정에 관한 연구)

  • Kim Sung;Noh Jae-Hyung;Kim Sung;Kim Yong-Il;Kim Jin-Pok
    • Journal of Gastric Cancer
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    • v.3 no.2
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    • pp.97-103
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    • 2003
  • Purpose: This study aims to determine the infection rate of Helicobacter pylori (H. pylori) in gastric cancer patients who received gastrectomies, and to compare the rates of H.pylori infection detected by serological test and that of histopathological test, and to evaluate its clinical meaning. Materials and Methods: Fifty two patients were selected from those who underwent gastrectomies at the Department of Surgery, Samsung Medical Center, from March 1997 to May 1997. The control group consisted of healthy 103 persons visited the center for health promotion in Samsung Medical Center. In both groups, we quantitatively checked serum level of IgG anti H. pylori antibody titer by ELISA using GAP IgG test kit (BioRad, USA) for the serological test, and we microscopically examined the surgical specimen stained by Warthin-Starry silver staining method for the histopathological test. Results: The seropositive rate of H. pylori in the patients' group was $71.2\%$ (37/52), and the control group was $65.0\%$ (67/103). The difference between two groups was statistically significant. However the histopathological study showed that the overall detection rate of H. pylori was $61.5\%$ (32/52) in the patients' group and $61.2\%$ (63/103) in the control group; nd this difference was not statistically significant Conclusion: We could confirm that H.pylori infection rate in the gastric cancer resected patients was statistically higher than in the normal healthy persons even in small population. And the detection method for the H. pylori infection by serological test was presumed to be better than that of histopathological test using surgical specimen. Further study for the larger population by well-organized multicenters will be needed.

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Generation of Antibodies Against Rice stripe virus Proteins Based on Recombinant Proteins and Synthetic Polypeptides

  • Lian, Sen;Jonson, Miranda Gilda;Cho, Won-Kyong;Choi, Hong-Soo;Je, Yeon-Ho;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.27 no.1
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    • pp.37-43
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    • 2011
  • Rice stripe virus (RSV) is one of serious epidemic pathogens for rice species grown in many Asian countries. Therefore, it is necessary to produce a diagnostic detection kit applicable in fields for RSV detection. In this study, RSV proteins that were derived from recombinant proteins and synthetic polypeptides as antigens were generated and were raised in rabbits for antiserum production. Among seven proteins in RSV, genes that code for NCP and NS3 proteins were cloned and subcloned into vector carrying His-tag protein and were expressed in E. coli. Of two recombinant proteins, only anti-NCP displayed stable hybridization signals in western blot analysis. Alternately, synthetic RSV polypeptides for CP, NCP, NS3 and NSvc4 we also generated and only antibodies against CP and NCP were very effective to detect RSV in both RSV infected rice and weed plants. However, antibodies against NS3 and NSvc4 showed weak specific bands as well as strong non-specific background due to the difference of viral proteins produced in the infected leaves. In summary, the antibodies generated against RSV proteins produced in this study will be useful for various assays such as for RSV diagnostic detection, immunoprecipitation, protein purification, and western blot analysis.