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1.
检测禽流感病毒抗体的重组核蛋白间接ELISA方法的建立 总被引：9，自引：3，他引：9
以大肠杆菌系统表达的H9N2亚型禽流感病毒（AIV）核蛋白（NP）为抗原，建立了禽流感间接酶联免疫吸附试验抗体检测技术（NP—ELISA）。对263份待检血清（包括临床收集的243份血清和20份H9N2亚型AIV免疫鸡阳性血清）进行检测，NP—ELISA与琼脂免疫扩散试验（AGP）的总符合率为83．3％，与血凝抑制试验（HI）的总符合率为92％。特异性试验表明，NP—ELISA方法可以检测H5、H7和H9亚型AIV特异性抗体，检测为阳性的血清样品能够被阳性鸡胚尿囊液阻断。敏感性试验证实，NP—ELISA最早可以检测鸡感染后7d的血清样品，并于感染后10d确定100％血清阳性，而AGP检测直到首免后21～28d才出现部分血清阳性，HI检测直到10～14d才出现部分血清阳性，并且NP-ELISA要比HI敏感4～40倍。试验证明，NP—ELISA是检测AIV血清型特异性抗体的一种特异、敏感、快速、经济的血清学检测技术。 相似文献
2.
以纯化的禽流感病毒（avian influenza virus,AIV）核蛋白作为捕捉抗原，金标兔抗鸡抗体作为金标二抗，建立检测禽流感血清抗体的胶体金免疫层析试纸条（immunochromatographic strip, ICS）。对标准阳性血清不同滴度进行检测，ICS试验的敏感性高于琼脂扩散试验（AGP）。特异性试验证实，禽流感的ICS试验与新城疫（ND）、传染性支气管炎（IBV）、鸡传染性法氏囊病（IBD）及减蛋综合症（EDS-76）均无交叉反应。同时，用ICS试验和HI试验检测105份鸡血清，两者具有较好的符合性。结果认为：禽流感ICS具有快速简便、特异、灵敏等优点。 相似文献
3.
本研究采用ＡＧＰ、ＨＩ等试验方法，对经Ｈ９亚型禽流感油乳剂灭活苗免疫、免疫后攻毒以及经Ｈ９Ｎ２活毒人工感染后的ＳＰＦ鸡抗体产生、消长规律进行了测定，结果表明人工感染ＳＰＦ鸡和免疫鸡一周后，ＡＧＰ的检出率即可达到１００％；Ｈ９亚型禽流感油乳剂灭活苗自免疫后一周内即可产生ＨＩ抗体，２１－２８天达到高峰，并能对相同亚型病毒感染引发良好的免疫反应。 相似文献
4.
Yamamoto T Tsutsui T Nishiguchi A Kobayashi S Tsukamoto K Saito T Mase M Okamatsu M 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(6):673-675
In June 2005, an outbreak of avian influenza (AI) caused by a low pathogenic H5N2 virus was identified in Japan. A serological surveillance was conducted because the infected chickens did not show any clinical signs. The Markov Chain Monte Carlo Method was used to evaluate the performances of serological HI and AGP tests because there was not enough time when the surveillance was initiated to conduct a test evaluation. The sensitivity of the AGP test (0.67) was lower than that of the HI test (0.99), while the specificities were high for both tests (0.96 for AGP and 0.90 for HI). Based on the low sensitivity of the AGP test, the HI test was used for primary screening in later stages of the epidemic. 相似文献
5.
The enzyme-linked immunosorbent assay (ELISA) and the conventional hemagglutination-inhibition (HI) test were compared for their ability to measure the primary serological response of chickens inoculated by the intranasal-intraocular (IN-IO) routes with Newcastle disease virus (NDV) and the secondary response after intratracheal (IT) challenge. In addition, these responses were compared with the temporal antibody response of chickens inoculated only once by the IT route. Both tests detected NDV-specific antibody by 7 days postinoculation (PI) in the IN-IO-inoculated group, while ELISA and the HI test detected antibody at 4 and 7 days, respectively, in the IT-inoculated group. Titers measured by each test were parallel in quantifying the antibody response, and titers rose anamnestically in response to secondary IT challenge at 21 days PI. ELISA titers remained high at 42 days PI, but the HI titers began to decline at this time. There was a good agreement (R = 0.94) between the results of the two tests throughout both primary and secondary responses. Conversely, there was little agreement between the results of the two tests after 21 days PI in the absence of secondary challenge. Antibody levels were higher when inoculation was by the primary IT route, and they persisted throughout the experiment (86 days). Ciliary activity served as a measure of tracheal immunity or infection of tracheal epithelium. It was reduced as early as 2 days PI and was nearly or completely absent by 5-6 days PI.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
6.
Effect of route of administration on the efficacy of a recombinant fowlpox virus against H5N2 avian influenza. 总被引：6，自引：0，他引：6
A recombinant fowlpox vaccine virus containing the H5 hemagglutinin gene of avian influenza virus was administered to susceptible chickens via wing-web puncture, eye drop, instillation into the nares, and drinking water. Even though there was a negligible hemagglutination-inhibition (HI) serologic response, all 10 chickens vaccinated by wing-web puncture remained without obvious signs of disease and survived challenge with a highly pathogenic strain of H5N2 avian influenza virus. All unvaccinated chickens and those vaccinated by nasal and drinking-water routes died following challenge. Eight of 10 chickens vaccinated with the recombinant by eyedrop died. All vaccinates were negative on the agar gel precipitin (AGP) test, and only one chicken had a positive HI titer before challenge. All chickens that survived challenge had high levels of HI antibody and were positive on the AGP test, indicating that they were infected by the challenge virus. 相似文献
7.
Avian influenza (AI) is a serious infectious disease caused by avian influenza virus (AIV) belonging to type A Orthomyxovirus. In the present study, we developed an indirect enzyme-linked immunosorbent assay (ELISA) employing E. coli-expressed full-length nucleoprotein (NP) of H9N2 avian influenza virus for the detection and quantification of antibodies
against AIV nucleoprotein. The NP-ELISA was compared with the AI agar gel propagation (AGP) test, haemagglutination inhibition
(HI) test, and IDEXX-FlockChek ELISA using 263 sera. The NP-ELISA was significantly more sensitive than the AGP and HI tests,
and showed 96.2% agreement ratio with IDEXX-FlockChek ELISA. With results obtained using the NP-ELISA, an ELISA titre (ET)
prediction equation, with which the ELISA titres of a flock or individual chickens can be determined, was derived from a positive/negative
(P/N) ratio standard curve. The NP-ELISA enables an alternative rapid serological diagnosis and is suitable for influenza
A antibody screening, especially in species that harbour several influenza subtypes. 相似文献
8.
为评价共表达鸡IL-6和H5亚型禽流感病毒HA基因重组鸡痘病毒(rFPV-AIH5AIL6)的免疫抗体消长规律及免疫效力,将重组病毒通过颈部皮下注射和翅部皮下注射2种不同的免疫途径来免疫鸡群,结果发现,2种免疫途径中,重组鸡痘病毒对鸡体质量增加均无影响,而野生型鸡痘病毒均可抑制鸡体质量增加。翅部皮下注射疫苗组能够产生较高的血凝抑制(HI)抗体水平,免疫21d后抗体水平达到高峰,28d后开始下降,49d时仍保持在一定的水平。免疫SPF鸡21d后攻毒,表明该重组病毒能使经滴鼻攻毒的SPF鸡抵抗H5亚型AIV的致死性攻击,保护率为95%,与油苗组相同,与单表达H5亚型禽流感病毒HA基因重组鸡痘病毒组(40%)差异显著;攻毒后3、5、7d采集喉头、泄殖腔棉拭子检测排毒情况,结果发现第3天排毒率最高,其中rFPV-AIH5AIL6免疫组排毒率为最低,显示IL-6在rFPV-AIH5IL6免疫过程中起到了免疫佐剂的作用,这为研制新型的禽流感重组鸡痘病毒疫苗奠定了基础。 相似文献
9.
Four- and six-week-old turkeys were vaccinated subcutaneously using avian influenza virus (AIV) A/Duck/613/MN/79 (H4N2) killed oil-emulsion vaccine. Sequential serological tests using agar gel precipitin (AGP), hemagglutination inhibition (HI), and enzyme-linked immunosorbent assay (ELISA) for measuring antibodies to AIV were performed up to 4 weeks postvaccination, when birds were challenged intranasally using A/Turkey/MN/80 (H4N2) live AIV. The ELISA was 25 to 1600 times more sensitive than the HI test and was able to detect antibody production earlier than the HI test. All turkeys with an ELISA titer of greater than or equal to 800 were protected against homologous challenge, as measured by virus recovery 3 days postchallenge. Four turkeys out of 20 serologically negative by AGP and HI tests but ELISA-positive were protected. 相似文献
10.
The immune response of individual chickens exposed in an aerosol apparatus to live commercial avian infectious bronchitis virus (IBV) vaccines was measured by serum neutralization (SN), haemagglutination inhibition (HI), complement fixation (CF) and agar gel precipitin (AGP) tests over a period of 14 weeks.The SN titres showed considerable variation for individual chickens and in a large number of birds were negative until 14 weeks after infection.Positive HI titres were recorded for most birds at one week after infection and persisted throughout the observation period. Some relationship was seen between HI and SN titres particularly in birds showing a high immune response.The results obtained with the AGP were transient, variable and did not compare well with results obtained by the other tests. The highest number of positive AGP reactors were seen two to three weeks after infection.Most birds showed positive titres by the CF test at some time after infection but titres were always low and did not correlate with results obtained by the other tests.Twenty-two weeks after vaccination 23 chickens were challenged by aerosol exposure to the Massachusetts 41 strain of IBV and four days later the trachea, kidneys and oviducts were removed from each bird for attempted virus isolation. Virus was recovered from only one kidney and 11 trachea samples. The mean pre-challenge HI and SN titres of birds from which no virus was recovered were significantly higher than the mash titres of vaccinated birds from which virus was isolated after challenge. 相似文献
11.
H9亚型禽流感病毒流行毒株交叉免疫攻毒保护试验 总被引：3，自引：0，他引：3
采用1998-2009年在河北、河南及山东分离的3株禽流感H9亚型流行毒株,分别制备灭活疫苗,免疫SPF鸡,免疫后21 d,采血测定HI抗体,然后用从上述3个地区及北京分离的共5株禽流感H9亚型流行毒株进行攻击,观察不同时期及地点分离的H9亚型流行毒株的交叉免疫攻毒保护效果。结果显示,用不同时期及地点的3个分离毒株所制备出的灭活疫苗免疫鸡后,各免疫组试验鸡H9亚型禽流感的HI抗体效价均明显上升,不同毒株灭活疫苗所诱导产生的HI抗体效价存在着不同程度的差异,用同源毒株作为抗原测定免疫组鸡的血清样品,可获得较高的HI抗体效价。攻毒试验结果证明,对不同时期及地点分离的禽流感H9亚型流行毒株间产生了较好的交叉保护力。用1998年分离的WD98株制备出的灭活疫苗对目前的流行毒株仍具有较好的保护效力。 相似文献
12.
In general, avian influenza (AI) vaccines protect chickens from morbidity and mortality and reduce, but do not completely prevent, replication of wild AI viruses in the respiratory and intestinal tracts of vaccinated chickens. Therefore, surveillance programs based on serological testing must be developed to differentiate vaccinated flocks infected with wild strains of AI virus from noninfected vaccinated flocks in order to evaluate the success of vaccination in a control program and allow continuation of national and international commerce of poultry and poultry products. In this study, chickens were immunized with a commercial recombinant fowlpox virus vaccine containing an H5 hemagglutinin gene from A/turkey/Ireland/83 (H5N8) avian influenza (AI) virus (rFP-H5) and evaluated for correlation of immunological response by hemagglutination inhibition (HI) or agar gel immunodiffusion (AGID) tests and determination of protection following challenge with a high pathogenicity AI (HPAI) virus. In two different trials, chickens immunized with the rFP-H5 vaccine did not develop AGID antibodies because the vaccine lacks AI nucleoprotein and matrix genes, but 0%-100% had HI antibodies, depending on the AI virus strain used in the HI test, the HI antigen inactivation procedure, and whether the birds had been preimmunized against fowlpox virus. The most consistent and highest HI titers were observed when using A/turkey/Ireland/83 (H5N8) HPAI virus strain as the beta-propiolactone (BPL)-inactivated HI test antigen, which matched the hemagglutinin gene insert in the rFP-H5 vaccine. In addition, higher HI titers were observed if ether or a combination of ether and BPL-inactivated virus was used in place of the BPL-inactivated virus. The rFP-H5 vaccinated chickens survived HPAI challenge and antibodies were detected by both AGID and HI tests. In conclusion, we demonstrated that the rFP-H5 vaccine allowed easy serological differentiation of infected from noninfected birds in vaccinated populations of chickens when using standard AGID and HI tests. 相似文献
13.
鸡抗REV血清的制备与检定 总被引：1，自引：1，他引：0
用网状内皮组织增生症病毒（REV）HA9901株免疫SPF鸡,无菌采血制备鸡抗REV血清。通过ELISA、IFA、AGP、HI等方法对制备的血清进行检测,未检出禽类常见的其他17种（或亚型）病毒的抗体,该血清具有良好的特异性。通过测定,该血清用于间接免疫荧光试验（IFA）的效价为1：2000,用于IFA检测REV的染色效果与REV单抗（11818和11854的混合物）相当。试验结果表明,该批血清1000倍稀释可作为一抗,用于REV的间接免疫荧光检测。 相似文献
14.
The H7N2 subtype of avian influenza virus (AIV) field isolate (H7N2/chicken/PA/3779-2/97), which caused the 1997-98 AIV outbreak in Pennsylvania, was evaluated for its infectivity, length of infection, and immune response in specific-pathogen-free (SPF) chickens. The composite findings of three clinical trials with various concentrations of virus indicated that this H7N2 subtype contained minimal pathogenicity for chickens. The concentration of the virus in the inoculum proved critical in the establishment of a productive infection in a chicken. Seven-day-old SPF chickens were not infected when inoculated with 10(0.7-2.0) mean embryo lethal dose (ELD50) of the H7N2 virus per bird. At this dose level, the immune response to this virus was not detected by the hemagglutination-inhibition (HI) test. Nonetheless, chickens at ages of 5 and 23 wk old tested were successfully infected when exposed to 10(4.7-5.7) ELD50 of H7N2 infectious doses per bird by various routes of administration and also by direct contact. Infected birds started shedding virus as early as 2 days postinoculation, and the period of virus shedding occurred mostly within 1 or 2 wk postinoculation (WPI). This H7N2 subtype of AIV induced a measurable immune response in all birds within 2 wk after virus exposure. Antibody titers were associated with AIV infectious doses and age of exposure of birds. Challenge of these infected birds with the same H7N2 virus at 5 and 10 WPI indicated the infective virus was recoverable from cloacal swabs at 3 days postchallenge and disappeared thereafter. In these challenged birds, the antibody levels as measured by the HI test spiked within 1-2 wk. 相似文献
15.
Prevalence and incidence of Newcastle disease and prevalence of Avian Influenza infection of scavenging village chickens in Timor-Lesté 总被引：1，自引：0，他引：1
Serrão E Meers J Pym R Copland R Eagles D Henning J 《Preventive veterinary medicine》2012,104(3-4):301-308
A longitudinal study to monitor prevalence and incidence of antibodies against Newcastle disease (ND) virus and prevalence of antibodies against Avian Influenza (AI) virus in scavenging village chickens was conducted in 20 villages within 4 districts of Timor-Lesté. A total of 3600 blood samples was collected from 1674 individual birds in 300 household chicken flocks during three sampling periods (December 2008-February 2009, March-May 2009, and June-August 2009). The mean interval between household visits was 101.6±1.9 days. None of the birds enrolled in the study was vaccinated against ND or AI. A haemagglutination inhibition (HI) test was used to determine antibody titres against ND virus and a competitive ELISA and HI tests were used to detect antibody against AI virus. The bird-level ND seroprevalence pooled across all samplings (adjusted for clustering by households) was 4.4% (95% CI 3.5-5.2). The bird-level ND seroprevalence in each of the three sampling periods (adjusted for clustering by household) was 3.0% (95% CI 2.0-4.0), 6.6% (95% CI 5.1-8.0) and 3.6 (95% CI 2.5-4.6), respectively. A total of 12.6% individual birds tested ND seropositive at least once over the total study period (95% CI 10.5-14.7). The flock-level ND seroprevalence (at least one bird tested had antibodies against ND virus) pooled across all samplings was 15.9% (95% CI 13.5-18.3). A total of 35.3% flocks had a minimum of one bird being ND seropositive at least once over the study period. The bird-level incidence rate for the period between the first and the second sampling and between the second and the third sampling was 5.6 (95% CI 4.1-7.5) and 0.5 (95% CI 0.5-3.8) per 10,000 bird-years-at-risk, respectively. A total of 1134 serum samples from the last sampling period between June and August 2009 was tested for antibodies against AI virus. Only 4 samples tested Influenza A positive, indicating a bird-level seroprevalence level for Influenza A of 0.4% (CI 0.0-0.7%). These Influenza A positive samples were further tested for HI antibodies against AI virus subtypes of H5N1, H5N3, H7N3 and H9N2, but all tested negative, suggesting that the influenza antibodies in those four birds resulted from exposure to low pathogenic AI viruses of different H subtypes. Our results indicate that village chickens in Timor-Lesté are exposed to ND virus; there was a higher risk of infection during the early months of 2009 than either immediately prior or subsequent to this. No evidence of infection of village chickens with H5, H7 or H9 AI viruses was detected in this study. 相似文献
16.
Dae Sub Song Youn Jeong Lee Ok Mi Jeong Yong Joo Kim Chan Hee Park Jung Eun Yoo Woo Jin Jeon Jun Hun Kwon Gun Woo Ha Bo Kyu Kang Chul Seung Lee Hye Kwon Kim Byeong Yeal Jung Jae Hong Kim Jin Sik Oh 《Journal of veterinary science (Suw?n-si, Korea)》2009,10(4):323-329
Active serologic surveillance is necessary to control the spread of the avian influenza virus (AIV). In this study, we evaluated a commercially-available cELISA in terms of its ability to detect AIV antibodies in the sera of 3,358 animals from twelve species. cELISA detected antibodies against reference H1- through H15-subtype AIV strains without cross reactivity. Furthermore, the cELISA was able to detect antibodies produced following a challenge of the AIV H9N2 subtype in chickens, or following vaccination of the AIV H9 or H5 subtypes in chickens, ducks and geese. Next, we tested the sensitivity and specificity of the cELISA with sera from twelve different animal species, and compared these results with those obtained by the hemagglutination-inhibition (HI) test, the "gold standard" in AIV sera surveillance, a second commercially-available cELISA (IZS ELISA), or the agar gel precipitation (AGP) test. Compared with the HI test, the sensitivities and specificities of cELISA were 95% and 96% in chicken, 86% and 88% in duck, 97% and 100% in turkey, 100% and 87% in goose, and 91% and 97% in swine, respectively. The sensitivities and specificities of the cELISA in this study were higher than those of IZS ELISA for the duck, turkey, goose, and grey partridge sera samples. The results of AGP test against duck and turkey sera also showed significant correlation with the results of cELISA (R-value >0.9). In terms of flock sensitivity, the cELISA correlated better with the HI test than with commercially-available indirect ELISAs, with 100% flock sensitivity. 相似文献
17.
免疫剂量和母源抗体对禽流感重组鸡痘病毒活载体疫苗免疫效力的影响 总被引：4，自引：0，他引：4
利用表达 H 5亚型禽流感病毒 (AIV)血凝素基因的重组鸡痘病毒 (r FPV- HA)以不同剂量免疫 1日龄 SPF鸡、有或无母源抗体 (FPV、AIV H5)的商品鸡 ,并于免疫后 2 1d利用同亚型 AIV通过肌肉注射进行致死性攻击 ,通过检测免疫后 HI抗体应答、比较攻毒后发病率和死亡率评价免疫剂量和母源抗体对 r FPV- HA免疫效力的影响。结果发现 ,免疫后 2 1d,15 %～ 2 0 %的 SPF鸡和无母源抗体商品鸡可检出 HI抗体 ,而含母源抗体商品鸡检测不到 HI抗体。利用H5亚型 AIV致死性攻击后 ,10 3～ 10 6 PFU的 r FPV- HA可保护 95 %～ 10 0 %的 SPF鸡和无母源抗体商品鸡抵御强毒攻击 ,使之免于发病和死亡 ;而不同剂量 r FPV- HA接种的含母源抗体商品鸡有 80 %～ 90 %发病和死亡。结果表明 ,在较宽的免疫剂量范围内 ,r FPV- HA对 SPF鸡和无母源抗体商品鸡可提供良好的保护 ,显示出一定的应用前景 ;母源抗体影响 r FPV- HA诱导的免疫应答 ,且提高免疫剂量亦不能克服其干扰作用 ,这提示在实际应用中需优化免疫程序 ,避免母源抗体干扰。 相似文献
18.
Changes in values of haemagglutination inhibition (HI) antibody titre, rectal temperature (RT) and total protein (TP) were determined for Shaver Brown chickens infected with Newcastle disease virus (NDV) Kudu 113. The infected chickens came down with Newcastle disease by day 3 post infection (PI). The major clinical signs were depression, greenish diarrhoea, paralysis of legs and wings, opisthotonus and torticolis. Mortality and morbidity were 52% and 1000%, respectively. There were haemorrhagic lesions in the wall of the intestine, proventricular mucosa and caecal tonsils. There were necrosis and mononuclear cell infiltration of the liver, kidney and spleen. There was a significant increase in daily mean HI antibody titres from days 3 to 9 PI. Similarly, significant rise in daily mean RTs were noticed in the infected chickens from days 1 to 13 PI. On the other hand, there was a decrease in daily mean TP concentrations of infected chickens, beginning from day 3 PI, and the lowest concentration of 2.60 +/- 0.15 g/dl was obtained by days 7 and 11 PI. The values of HI, RT and TP for the control chickens were relatively constant during the experiment. The correlation coefficient (r) between HI and RT was positive and highly significant (r = 0.725, p<0.001), while the relationship between HI and TP was negative but highly significant (r = -0.712, p<0.001). It was concluded that NDV Kudu 113 induced increases in values of HI and RT, which occurred concurrently with a decrease in TP concentrations of infected chickens. 相似文献
19.
20.
Raj GD Thangavelu A Elankumaran S Koteeswaran A 《Tropical animal health and production》2000,32(3):173-178
The rocket immunoelectrophoresis (RIE) test was used for the qualitative detection and quantitative estimation of infectious bursal disease virus (IBDV) specific antigen in experimentally infected chickens and samples collected from suspected outbreaks. The IBDV specific antigen was detected in the bursae of experimentally inoculated chickens up to 5 days post infection (PI) by the agar gel precipitation (AGP) test and 7 days PI by the RIE test. The RIE detected IBDV specific antigen in a significantly greater number of samples collected from the field outbreaks than the conventional AGP test. Exudative bursae were found to have a higher antigen content than haemorrhagic bursae and are recommended as the material of choice for diagnosis of IBD. This test could also be used to quantify IBDV specific antigen in commercial killed vaccines. 相似文献