• Title/Summary/Keyword: serum antibody

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Rapid Detection of Serum HCV RNA by Combining Reverse Transcription and PCR without RNA Extraction

  • Jang, Jeong-Su;Lee, Kong-Joo
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.486-489
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    • 1996
  • A simple, rapid, specific and sensitive method for the detection of serum hepatitis C virus (HCV) RNA using the reverse transcription-polymerase chain reaction (RT-PCR) technique without conventional RNA extraction was developed. HCV template RNA from serum was obtained by boiling the serum at $95^{\circ}C$ for 2 min, cooling rapidly in ice and removing the proteins by cetrifugation. RT-PCR amplifications including the reverse transcription and first PCR amplification were performed in one vessel containing both of reverse transcriptase and Taq DNA polymerase. The detection of HCV RNA from $10^{-3}{\mu}l$. serum was possible with this method. The suitability of this method for clinical analysis was evaluated by assaying HCV RNA in 225 patient samples including anti-HCV antibody negatives (13 samples) and positives (212 samples) by enzyme-linked immunosorbent assay test (ELISA). Detections of HCV RNA with this method were in 4 of 13 anti-HCV antibody negative samples (30.8%) and 95 of 212 positive samples (44.8%). The present method can be completed in 1 hr and has a wide range of application for the clinical utilities to determine the viral RNAS.

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Studies on enzyme-linked immunosorbent assay(ELISA) for detection of antibody to Brucella abortus (효소면역법을 이용한 Brucella abortus 항체 검출에 관한 연구)

  • 심항섭;국정희;정봉수;고태오;조중현;박유순
    • Korean Journal of Veterinary Service
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    • v.21 no.2
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    • pp.107-115
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    • 1998
  • In order to establish a rapid, sensitive and specific diagnostic method for detection of antibody to Brucella abortus, a enzyme-linked immunosorbent assay(ELISA) was adapted. The diagnostic efficacy of the established ELISA was compared with that of the standard tube agglutination test for B abortus. 1. It was found that the optimal concentration of antigen for this ELISA was 5$\mu\textrm{g}$/ml, the optimal dilution of conjugate was 1 : 2000, and the optimal dilution of serum was 1 : 200, respectively. 2. Cut off value in this ELISA was 1,102 that was determined by mean absorbance(at 492nm) of tube agglutination test negative serum added with the triple value of the standared devation. 3. The relationship between the tube agglutination test and ELISA was showen high corresponding rate with sensitivity(96.3%) and specificity(98.1%). 4. The efficacy of the ELISA for detection of B abortus antibody was compared with tube agglutination test In brucellosis outbreak farm. The sensivity of ELSIA was higher than tube agglutination test.

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Prevalence of Leptospiral Antibodies in Pigs (돼지의 Leptospira 속균에 대한 혈중항체조사 -제천시 소재 도축장 출하돈 중심-)

  • 강신석;곽학구;김홍기
    • Korean Journal of Veterinary Service
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    • v.15 no.1
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    • pp.46-50
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    • 1992
  • This experiment was to investigate the Leptospiral antibody in the pigs with the serological test by the microscopic-agglutination-test (MAT) from November 1991. to Januaury 1992. Antigen(living antigen) was used L. icterohemorrhagiae, L. pomona, L. canicola, L. hardjo, L. ballum, L. australis, L. autumnalis, L. grippotyphosa, L. tarassovi, L. pyrogenes, L. bataviae, L. hebdomadis for the serolgical test in the pigs. The result obtained are summarized as follows of the total 202 serum samples examined, 1. Among the serum samples of 202 heads, 19 heads of the pigs(9.4%) were positive. 2. Among the positive samples of 19 heads, The detected were L. icterohemorhagiae 10 heads(5.0%), L. pomona 3 heads(1.5%), L. canicola 6 heads(3.0%). 3. Antibody titers of positive sera were ranging from 1:100 to 1 : 400. Serotiters appeared to be very low, 4. The seroprevalence of Leptospira in Chechon was higher than that other districts (5. 4% -5.8%), but the lower than Chung-nam, Kyonggi(13.7-15.9%)

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Radioimmunoassay of High-Density Lipoprotein in Japanese Quail Serum (메추리혈청 리포단백질의 방사면역분석법)

  • 한병훈;한용남;유시용;양현옥;박명환
    • YAKHAK HOEJI
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    • v.29 no.2
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    • pp.90-95
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    • 1985
  • The development of radioimmunoassay (RIA) for high-density lipoprotein (HDL) in Japanese quail serum will contribute to the screening of drugs acting on cholesterol transport. We have developed a double antibody RIA method for J. quail HDL. The first antibody was raised in rabbit by immunization of HDL isolated by the dextrane sulfate-$Mn^{#}$ precipitation method. For the preparation of raclioiodinated antigen, HDL was further purified by combination of electrophoretic procedure. Using the second antibody raised in goat by rabbit IgG, we have furnished the RIA method for HDL. It showed high specificity and sensitivity of working assay range, 0.1-33.mu.g HDL/tube. There was no correlation between the radioimmunoassay of HDL and the enzyme assay of HDL-cholesterol.

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Antifertility Effect of Passive Immunization against Progesterone Monoclonal Antibody in Mice (Progesterone 단일클론항체의 수동면역이 Mouse 수정란의 착상저해에 미치는 효과)

  • 김정우;김종배;정길생;고대환
    • Korean Journal of Animal Reproduction
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    • v.14 no.3
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    • pp.157-164
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    • 1990
  • Anti-progesterone monoclonal antibody injected intraperitonially as a single dose(100$\mu\textrm{g}$) 48hours post coitum(p.c.) almostly blocked pregnancy in ICR mice. The blocking rate of pregnancy in mice treated with antibody were decreased proportionally according to dose of antibody injected ; the rate were 60%, 57% and 17% as the antibody of 10$\mu\textrm{g}$, 50$\mu\textrm{g}$ and 100$\mu\textrm{g}$ were injected respectively. Blood serum progesterone concentration was greatly increased(21 times) after treatment(100$\mu\textrm{g}$), by virtue of high-affinity binding by antibody in circulation of non-pregnant mice in coompared with that of control group at day 10 p.c.. The concentration was about 1.6 times higher in the pregnant mice than in the non-pregnant mice in antibody treated group. In control group, the progesterone concentration was over 7 times higher in the pregnant mice than in non-pregnant mice at day 5 p.c..

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AN INVESTIGATION OF IMMUNIZATION AGAINST SOMATOSTATIN BY MEASURING ANTIBODY TITRES, SOMATOSTATIN AND SOMATOTROPIN PROFILES IN GILTS

  • Du, Z.L.;Hacker, R.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.2
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    • pp.211-218
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    • 1993
  • The effect of active immunization against porcine somatostatin (SRIF-14) on somatostation and somatotropin secretion profile in 18 gilts was investigated. Gilts were assigned to the following treatments: control (sham injection, n = 6); bovine serum albumin (BSA) (injection of BSA with bacterial protein adjuvant, n = 6); SRIF (injection of BSA-SRIF-14 conjugate with bacterial protein adjuvant n = 6). Serum SRIF and pST were assayed from the blood samples taken on day 7 after the last immunization injection. Anti-SRIF antibody titres were assayed in weekly samples two weeks after the initial immunization to one week after the last immunization. Results revealed that the immunization protocol used in the present investigation failed to produce antibodies capable of neutralizing endogenous somatostatin. In addition, the porcine somatotropin assay revealed no significant differences in baseline pST concentration, mean peak amplitude and number of peaks during a 24 h secretory period among SRIF, BSA and control treatment. There were also no differences in SRIF baseline concentration, peak amplitude, and number of peaks during a 24 h secretory period among any of the three treatments. Circulating concentrations of pST and pSRIF were highly correlated (r = -0.09). Furthermore, anti-SRIF antibody titre was not detected in the serum of the gilts actively immunized against SRIF. These data, collectively, suggest that the protocol employed in the present investigation for active immunization against SRIF is not an effective method for changing SRIF and pST secretion profiles of the gilt and thus to enhance performance.

Rapid Detection of Ovarian Cancer from Immunized Serum Using a Quartz Crystal Microbalance Immunosensor

  • Chen, Yan;Huang, Xian-He;Shi, Hua-Shan;Mu, Bo;Lv, Qun
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3423-3426
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    • 2012
  • Background: The objective of this study was to measure the antibody content of NuTu-19 ovarian cancer cells in serum samples using a quartz crystal microbalance (QCM) immunosensor. Materials and Methods: NuTu-19 cells were first cultured onto the electrode surfaces of crystals in Dulbecco's modified Eagle medium, and then specified amounts of immunized serum samples of immunized rabbit were also added. The change in mass caused by specific adsorbtion of antibodies of NuTu-19 to the surfaces of the crystals was detected. Results: The change in resonance frequency of crystals caused by immobilization of NuTu-19 cells was from 83 to 429Hz. The antibody content of NuTu-19 detected was 341ng/ul. The frequency shifts were linearly dependent on the amount of antibody mass in the range of 69 to 340ng. The positive detection rate and the negative detection rate were 80% and 100%, respectively. Conclusion: This immunoassay provides a viable alternative to other early ovarian cancer detection methods and is particularly suited for health screening of the general population.

Distribution of the serum Ig G titers to whole cell and leukotoxin of Mannheimia haemolytica Al in Holstein and Korean indigenous cattle slaughtered in abattoir

  • Byun, Jae-Won;Kim, Kyung-Ho;Lee, Sung-Mo;Lee, Jung-In;Hwang, Hyun-Soon;Kim, Yong-Hee
    • Korean Journal of Veterinary Service
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    • v.26 no.4
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    • pp.323-328
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    • 2003
  • A total of 419 slaughtered cattle were used to investigate the serum Ig G titers to the Mannheimia haemolytica Al whole cell and leukotoxin recognized with important virulence factor in bacterial pathogenesis. Data obtained in this study were represented with average absorbance${\pm}$standard deviation. Serum Ig G titers were detected with the ranges from 0.1 to 0.5 at 490nm. Whole cell titers were higher than leukotoxin antibody on the whole. Antibody titers of slaughtered cattle between races, ages have no significant difference but gradual decrease under aging in dairy cow for whole cell (decline mean titer from 0.29 to 0.27 according to age) was undertaken. Holstein bulls shipped from Seoul province had a significantly lower Ig G titers than those from another ones (p<0.05).

Effect of Serum Type on Hybridoma Growth and Monoclonal Antibody Production (하이브리도마 세포증식과 단일클론항체 생산에 미치는 혈청 종류의 영향)

  • 전복환;박송용
    • KSBB Journal
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    • v.9 no.3
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    • pp.253-265
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    • 1994
  • We have studied the effects of serum concentration and initial cell density on hybridoma cell growth and monoclonal antibody (MAb) production at various media supplemented with different types of serum. The types of serum were fetal bovine sera, newborn bovine calf sera, calf sera including supplemented calf sera, horse serum, and goat serum. The concentrations of each serum were 0.5, 1.25, 2.5, and 5% (v/v) and the inoculum densities were $5{\times}10^4, 1{\times}10^5, 2{\times}10^5,$ cells/ml. The hybridoma cell growth and anti-Hepatitis B surface antigen (anti-HBsAg) MAb production were found to be enhanced by increasing the serum concentration and by increasing inoculum density regardless of serum type. We found that test sera purchased from different companies show different effects on cell growth and MAb production, although they are the same type of serum.

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Production and Characterization of DDT Antibodies and Its Application to Enzyme Immunoassay: Relation of Response and Affinity to Coating Ligand

  • Hong, Ji-Youn;Kim, Jong-Hyun;Park, Song-Ja;Lho, Dong-Seok;Choi, Myung-Ja
    • Bulletin of the Korean Chemical Society
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    • v.24 no.11
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    • pp.1605-1608
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    • 2003
  • To development an immunodetection method for DDT, 1,1,1-trichloro-2,2-bis(4-chorophenyl)ethane (p,p'-DDT) and its metabolites (p,p'-DDA, p,p'-DDE, p,p'-DDD), five derivatives of DDT haptens have been synthesised and characterized as coating ligands for antibody evaluation. The appropriate lengths of linkers were introduced to investigate a matching pair of coating ligand and antibody. Among these hapten derivatives, 2,2-bis(4-chlorophenyl)acetic acid (DDA), 5,5-bis(4-chlorophenyl)-5-hydroxypentanoic acid (DDHP) and 5,5-bis(4-chlorophenyl)-5-chloropentanoic acid (DDCP) were conjugated with keyhole limpet hemocyanin (KLH) for its use as an immunogen. The bovine serum albumin (BSA) conjugates of these derivatives were prepared as a coating ligand for monoclonal antibody screening. Fifteen monoclonal antibody clones were screened using these probes. 6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoic acid (DDHH) and 3-[6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoylamino]propanoic acid (DDHHAP), in addition to the above hapten derivatives, were conjugated to ovualbumin (OVA) and bovine serum albumin (BSA) for their use as coating ligands to measure the titration level of the antibody and the displacement of free analytes. The indirect competitive ELISA results indicate that the titration level and free analyte displacement were greatly influenced by the DDT derivatives and carrier proteins used. Three matching pairs of monoclonal antibodies and coating ligands were selected for the DDT immunoassay: antibody clone 1A3 and coating ligand DDA-OVA, 1A1 and DDHHAP-BSA, and 1A4 and DDHP-OVA.