• Title, Summary, Keyword: serological detection

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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.

Comparative study of serological detection methods on old bloodstain samples (오래된 혈흔 시료에 대한 혈청학적 식별법의 비교 연구)

  • Lee, Minji;Ahn, Eu-Ree;Kim, Da-Hye;Shin, Heejin;Jung, Ju Yeon;Lee, Sung-Jin;Chun, Byung-Won
    • Analytical Science and Technology
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    • v.31 no.5
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    • pp.201-207
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    • 2018
  • Blood is a commonly found body fluid at crime scenes, and plays an important role in identifying suspects and in the reconstruction of crime scenes. Although serological detection of blood has been widely used in the field of forensic science, research on the detection of old bloodstains is scarce. This work aimed to compare various methods for the detection of old bloodstains and validate the reliability of their results. Four presumptive tests-Tetramethylbenzidine, $Bluestar^{(R)}$, Leucomalachite Green, Kastle-Meyer tests-and two confirmatory tests-Fecal Occult Blood (FOB) and Rapid Stain $Identification^{(TM)}-Blood$ ($RSID^{TM}-Blood$) tests-were compared. Bloodstain samples from post-mortem cases were collected on gauzes and then stored at room temperature for periods from 7 to 30 years. All the presumptive tests were positive, even for the 30-year-old sample. However, FOB and $RSID^{TM}-Blood$ provided false negative results for some samples stored for 17 years or more (1988 to 2001). The results indicate that FOB and $RSID^{TM}-Blood$ are not reliable for the detection of old bloodstains. These findings can be useful in the selection of an appropriate detection method for serological testing of old bloodstains. In addition, the information will be useful background knowledge when applied in the field of forensic practice.

Pathological and serological detection of bovine viral leukosis in a dairy farm in Jeonbuk province (유우농장에서 발생한 소바이러스성 백혈병의 병리학적 및 혈청학적 조사)

  • Jo Young-Suk;Jang Sae-Gun;Chu Keum-Suk;Choi Eun-Young;Chon Hee-Woong;Hong Jae-Hee;Lim Chae-Woong
    • Korean Journal of Veterinary Service
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    • v.29 no.2
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    • pp.89-96
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    • 2006
  • Bovine viral leukosis is a viral disease of cattle characterized by the development of tumors in the lymphatic tissue. A female Holstein, 3-year-old, was submitted for diagnosis at the Diagnostic laboratory, Chonbuk National University. Clinical sign of the affected animal showed emaciation, enlargement of superficial lymph node and mild diarrhea. Remarkable lesions were enlargement of many internal lymph nodes. Histopathology revealed excessive neoplastic lymphoid cells characteristic of BVL infection. Subsequently, serums from all cattle were collected and serological examination was done where a 85% seropositive rate was detected using ELISA test. ELISA method showed a comparatively 75% higher detection rate than the agar gel immunodiffusion (AGID) test (85% vs 40%). Serologically positive cattle were variably detected in all ages from under 1 year to over 6 year of age. Hematological examination consistently showed leukocytosis and a differential lymphocytosis of seropositive cattle. Detailed comparative pathological and serological data diagnosed the presence of bovine viral leukosis.

Antiserum Preparation of Recombinant Sweet Potato Latent Virus-Lotus (SPLV-Lotus) Coat Protein and Application for Virus-Infected Lotus Plant Detection

  • He, Zhen;Dong, Tingting;Chen, Wen;Wang, Tielin;Gan, Haifeng;Li, LiangJun
    • The Plant Pathology Journal
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    • v.36 no.6
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    • pp.651-657
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    • 2020
  • Lotus is one of the most important aquatic vegetables in China. Previously, we detected sweet potato latent virus from lotus (SPLV-lotus) and found that it has highly significant sequence diversity with SPLV-sweet potato isolates (SPLV-sp). Here, we developed serological methods for the detection of SPLV-lotus in Chinese lotus cultivation areas. Based on the high sensitivity of SPLV-lotus coat protein antiserum, rapid, sensitive and large-scale diagnosis methods of enzyme-linked immunosorbent assay (ELISA) and dot blot in lotus planting area were developed. The established ELISA and dot blot diagnostic methods can be used to detect SPLV-lotus from samples successfully. And our results also showed that the SPLV-lotus and sweet potato isolates appeared clearly distinction in serology. Our study provides a high-throughput, sensitive, and rapid diagnostic method based on serology that can detect SPLV on lotus, which is suggested to be included in viral disease management approach due to its good detection level.

Noninvasive fetal RHD genotyping using cell-free fetal DNA incorporating fetal RASSF1A marker in RhD-negative pregnant women in Korea

  • Han, Sung-Hee;Yang, Young-Ho;Ryu, Jae-Song;Kim, Young-Jin;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.12 no.2
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    • pp.100-108
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    • 2015
  • Purpose: Conventional methods for the prenatal detection of fetal RhD status involve invasive procedures such as fetal blood sampling and amniocentesis. The identification of cell-free fetal DNA (cffDNA) in maternal plasma creates the possibility of determining fetal RhD status by analyzing maternal plasma DNA. However, some technical problems still exist, especially the lack of a positive control marker for the presence of fetal DNA. Therefore, we assessed the feasibility and accuracy of fetal RHD genotyping incorporating the RASSF1A epigenetic fetal DNA marker from cffDNA in the maternal plasma of RhD-negative pregnant women in Korea. Materials and Methods: We analyzed maternal plasma from 41 pregnant women identified as RhD-negative by serological testing. Multiplex real-time PCR was performed by amplifying RHD exons 5 and 7 and the SRY gene, with RASSF1A being used as a gender-independent fetal epigenetic marker. The results were compared with those obtained by postnatal serological analysis of cord blood and gender identification. Results: Among the 41 fetuses, 37 were RhD-positive and 4 were RhD-negative according to the serological analysis of cord blood. There was 100% concordance between fetal RHD genotyping and serological cord blood results. Detection of the RASSF1A gene verified the presence of cffDNA, and the fetal SRY status was correctly detected in all 41 cases. Conclusion: Noninvasive fetal RHD genotyping with cffDNA incorporating RASSF1A is a feasible, reliable, and accurate method of determining fetal RhD status. It is an alternative to amniocentesis for the management of RhD-negative women and reduces the need for unnecessary RhIG prophylaxis.

Sensitivity analysis of serological tests for detection of disease in cattle (소 질병 검출을 위한 혈청학적 검사의 민감도 평가)

  • Lee, Sang-Jin;Moon, Oun-Kyong;Pak, Son-Il
    • Korean Journal of Veterinary Research
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    • v.50 no.1
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    • pp.43-48
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    • 2010
  • Animal disease surveillance system, defined as the continuous investigation of a given population to detect the occurrence of disease or infection for control purposes, has been key roles to assess the health status of an animal population and, more recently, in international trade of animal and animal products with regard to risk assessment. Especially, for a system aiming to determine whether or not a disease is present in a population sensitivity of the system should be maintained high enough not to miss an infected animal. Therefore, when planning the implementation of surveillance system a number of factors that affecting surveillance sensitivity should be taken into account. Of these parameters sample size is of important, and different approaches are used to calculate sample size, usually depending on the objective of surveillance systems. The purpose of this study was to evaluate the sensitivity of the current national serological surveillance programs for four selected bovine diseases assuming a specified sampling plan, to examine factors affecting the probability of detection, and to provide sample sizes required for achieving surveillance goal of detecting at least an infection in a given population. Our results showed that, for example, detecting low level of prevalence (0.2% for bovine tuberculosis) requires selection of all animals per typical Korean cattle farm (n = 17), and thus risk-based target surveillance for high risk groups can be an alternative strategy to increase sensitivity while not increasing overall sampling efforts. The minimum sample size required for detecting at least one positive animal was sharply increased as the disease prevalence is low. More importantly, high reliability of prevalence estimation was expected with increased sampling fraction even when zero-infected animal was identified. The effect of sample size is also discussed in terms of the maximum prevalence when zero-infected animals were identified and on the probability of failure to detect an infection. We suggest that for many serological surveillance systems, diagnostic performance of the testing method, sample size, prevalence, population size, and statistical confidence need to be considered to correctly interpret results of the system.

The serological studies on infectious bursal disease (전염성 F낭병에 대한 혈청학적 연구)

  • 정영미;서석열;도홍기;조정곤;노수일
    • Korean Journal of Veterinary Service
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    • v.23 no.3
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    • pp.271-279
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    • 2000
  • This study was carried out to provide the fundamental information for development of proper vaccination program against infectious bursal disease(IBD) to the local chicken farms. The antigen detection was peformed from 8 samples of bursa of Fabricius with agar gel precipitation(AGP) and indirect immunofluorescent assay(IFA), And also, the antibodies in serum samples were detected by the various serological methods such as commercial ELISA assay, AGP and virus neutralization(VN) test. 1. The antigen detection rates were 25% for AGP which is 2 out of 8 farms and 10 out of 40 bursas, and 25% which Is 2 out of 8 farms and 20% 8 out of 40 bursas for IFA, respectively. 2. The mean titer of maternal antibody (>3,000) existed until 10 days of the age with ELISA-GMT. 3. The antibody positive rates which are over 80% showed until 5 days of the age with ELISA and at 10 days of the age with AGP except one, but none of them showed from 1 day of the age. This report came to conclusions that both the protective maternal antibody titers and the antigen positive rates were significant until at the 10 days of the age.

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Comparison of serological methods for detection of avian influenza virus antibodies (가금인플루엔자 바이러스 항체검출을 위한 혈청학적 진단법 비교)

  • Han, Myung-guk;Park, Kyoung-yoon;Kwon, Yong-kuk;Kim, Jae-hong
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.73-80
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    • 2002
  • An enzyme-linked immunosorbent assay (ELISA) using purified hemagglutinin of swine influenza virus (H1N1) as antigen was developed for detection of antibody to avian influenza virus (AIV). The sensitivity and specificity of a developed and commercial available ELISA kits were compared with those of agar gel precipitation (AGP) test and hemagglutination inhibition (HI) test using sera collected from chickens under condition of field exposure. The concentration of antigen, serum dilution and concentration of enzyme-conjugated secondary antibody in developed ELISA (S-ELISA) were 0.5ug/100ul, 1:200 and 0.03ug/100ul, respectively. The correlation coefficients between S-ELISA and commercial ELISA and HI titers were 0.419 and 0.533, respectively. A significant correlation (p < 0.01) was not found between HI and ELISA titers. The S-ELISA was found to be as more sensitive and specific than the AGP test, showing 86.8% sensitivity and 85.3% specificity. It is suggested that the ELISA using the SIV as antigen may be useful method as an investigating tool for AIV serological surveillance.

Comparison of Probability of Detecting Bluetongue in Quarantine Testing for the Imported Cattle with Special Focus on the Sampling Scenario (수입 소의 검역검사 수준에 따른 블루텅 검출 확률 비교)

  • Pak, Son-Il
    • Journal of Veterinary Clinics
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    • v.27 no.4
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    • pp.421-426
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    • 2010
  • In view of free from bluetongue (BT) in the domestic cattle population in Korea, the key of quarantine testing for BT virus (BTV) infection is detection of cattle previously exposed to the virus. The objective of this study was to estimate the probability of detecting a cattle infected with BTV using a stochastic modeling analysis of existing quarantine testing data. Three testing scenarios were considered in this study: serological testing of all animals in all imported lots (scenario 1), serological testing of a sample of cattle from all imported lots (scenario 2), and serological testing of 50% of imported lots (scenario 3). In scenario 2 and 3, it was assumed that cattle were sampled (sample size) within each lot to detect 5% of the cattle in each lot with a 95% confidence, taking into account diagnostic sensitivity of the ELISA (enzyme-linked immunosorbent assay). The model output was the total number of BTV-infected cattle and the prevalence of BTV infection in imported cattle from the US, Australia, Canada and Japan. Compared to the scenario 1, the probability of detecting a BTV-infected cattle was estimated to be 19% and 1.6% in scenario 2 and 3, respectively. Furthermore, the analyses showed a 95% confidence that BTV prevalence was less or equal to $9.7{\times}10^{-4}$ (median = $1.5{\times}10^{-5}$), indicating that, for the scenario 2 and 3 with serological testing for a sample of cattle, the risk of introducing an exotic strain of BTV into Korea through the importation of live cattle would not be acceptable.

Molecular fingerprinting of olive flounder pathogenic Streptococcus parauberis strains by random amplified polymorphic DNA analysis

  • Jung, Yong-Uk;Kang, Sang-Hyuck;Jin, Chang-Nam;Kang, Bong-Jo;Heo, Moon-Soo
    • 한국생물공학회:학술대회논문집
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    • pp.899-903
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    • 2005
  • Two infectious species of Streptococcosis pathogens were detected by multiplex PCR assay. Detection rates of Streptococcus iniae and S. parauberis could reach 44.9% and 55.1% respectively for one year during 2004 to 2005 in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeiu island. In the present study we have investigated the interspecific relationship of all Jeju area of S. parauberis by RAPD analysis. Represent strains divided to four groups by RAPD fingerprints. The important differences observed between the olive flounder isolates suggest that they could constitute a well-differentiated group or a separate clonal line within this bacterial species. Though, serological research of S. parauberis strains in Jeju island not exist yet. These strains doing the serological evolution.

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