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1.
支原体是生物制品中常见的一种污染微生物,工作环境、操作者本身、培养基、污染支原体的细胞、试验器材和用来制备细胞的原始组织或器官都可能给生产带来支原体的污染。生物制品一旦污染支原体,会引起质量下降,免疫效果得不到保证,严重时可造成人力、物力、财力上的巨大损失,因此在生物制品生产中,需严格保持无菌操作,建立快速有效的检测方法,有效防止支原体的污染。  相似文献   

2.
兽用生物制品是一类特殊的动物保健品,是控制和消灭动物传染病,保障动物及人类健康的重要武器之一。若兽用生物制品被支原体污染,制品的质量和安全将会受到严重影响,同时会造成巨大的经济损失。结合生产实际对兽用生物制品支原体污染的主要来源和支原体污染的检测方法进行了介绍,制定了科学合理的预防控制措施,以期为确保兽用生物制品无支原体污染,质量安全可靠提供参考。  相似文献   

3.
猪鼻支原体(Mycoplasma hyorhinis,Mhr)是引发猪地方性肺炎及仔猪多发性浆膜炎、关节炎的重要病原之一,也会引起猪场的常见消耗性疾病。Mhr常常污染细胞培养物,对疫苗生产造成巨大危害。定期对猪场常用的生物制品进行Mhr检测,可减少猪场由于污染的生物制品而造成损失。  相似文献   

4.
自1956年国外学者第一次检测到细胞培养物中存在支原体污染以来,这个世界性问题持续了半个多世纪。对于生物制品中支原体污染的检测方法多种多样,但尚无统一的内控标准,而建立一种快速、准确的方法用于支原体污染的检测,可以有效地减少和预防支原体污染,从而提高生物制品的纯净性。本文对支原体污染的几种常见检测及鉴定方法进行综述,包括DNA荧光染色法、PCR、IFA与ELISA等,以期为支原体污染的检测及鉴定研究提供参考。  相似文献   

5.
支原体对细胞培养污染的研究概况   总被引:1,自引:0,他引:1  
支原体是自然界中存在的最小、最简单的原核生物,大小介于细菌和病毒之间,可通过滤菌器,对许多抗生素有抗性,是细胞培养污染中一种常见的微生物.被支原体污染后的细胞培养物,引起细胞形态学和功能的改变,导致用细胞基质制备的生物制品报废,且很难清除.一旦发生污染,就意味着试验的失败,造成巨大的人力、物力、财力的浪费.从支原体对细胞培养物污染后造成的危害及对支原体污染的预防、检测、清除等的研究进展进行论述,为细胞支原体污染的防控提供依据和手段.  相似文献   

6.
为了解兽用生物制品支原体污染情况,对2007-2011年全国26家兽用生物制品企业生产的22个品种141批生物制品的支原体检验监督抽检情况进行了汇总,就存在的问题进行了分析,并从合理制订监督抽检计划、完善信息监督机制、提高产品质量、开展新方法研究等四个方面提出了建议,为下一步的兽用生物制品的监督抽检工作提供参考。  相似文献   

7.
为建立一种快速检测兽用生物制品中支原体污染的方法,试验根据GenBank上登录的支原体16S rRNA种属保守区序列设计1对引物,经PCR条件优化,建立了检测兽用生物制品中支原体污染的PCR方法。结果表明:该方法对大肠杆菌、沙门氏杆菌、巴氏杆菌、猪链球菌、猪圆环病毒2型、猪伪狂犬病毒、猪瘟病毒、猪繁殖与呼吸综合征病毒、新城疫病毒的扩增结果均为阴性;对鸡毒支原体、猪肺炎支原体、禽滑液支原体基因组DNA的检测灵敏度分别达到0.13,0.88,0.14 ng,与培养法的符合率达98.84%;应用该方法对鸡胚、血清、细胞等140份临床样品进行支原体污染的检测,阳性污染率达6.43%。表明PCR方法检测具有良好的特异性、敏感性、准确性和适用性,可用于兽用生物制品生产中鸡胚、血清、细胞、疫苗半成品等的质量控制和疫苗的质量检验。  相似文献   

8.
生物制品中的支原体污染   总被引:1,自引:0,他引:1  
<正>支原体(Mycoplasma)是一种大小介于细菌和病毒之间(最小直径0.2μm)并独立生活的微生物,是生物制品细胞培养中较常见的污染物。目前,细胞培养(特别是传代细胞)被支原体污染已成为一个世界性问题。据报道,目前世界上有30%~50%的细胞系(株)已受支原体污染。但是在生物制品制作过程中即使有大量的支原体污染,也不会像细菌、真菌那样在普通培养基上明显地观察到,必须有适宜其生长的培养基进行培养时才能发现。污染的支原体在组织培养中繁殖有  相似文献   

9.
兽用生物制品可以对动物传染病进行控制或灭除,有利于保障动物的健康以及人类的健康,属于一种类型较为特殊的保健品.但是如果支原体污染兽用生物制品,将能够导致兽用生物制品的使用效果及安全性受到不良影响,还有可能导致严重的经济损失,所以有必要根据实际生产情况,针对兽用生物制品之中支原体的检测方法及主要来源进行分析,并制定与实际...  相似文献   

10.
经微生物促生长及灵敏度检测合格的无菌及支原体检验培养基各3批,分别在6个兽用生物制品企业进行了应用试验。抽检疫苗52批和半成品抗原5批,3批无菌检验培养基检测结果一致,疫苗2/52批、半成品抗原3/5批污染。禽源支原体检验培养基抽检样品17批、非禽源支原体检验培养基抽检33批及血清支原体检验培养基抽检10批,3批培养基检测结果一致,均无支原体污染。  相似文献   

11.
在动物疫苗研制过程中,应用巢式PCR方法,设计两套引物,扩增支原体16S与23SrRNA基因间隔区序列,可以检测造成细胞污染的常见支原体种类。本试验应用该方法检测三批样品和阴性对照均无特异性目标条带,即三批疫苗样品支原体检测均为阴性。阳性对照样品在200-400bp之间出现特异性目标条带,实验表明所建立的巢式PCIL方法是一种快捷、灵敏、准确的检测方法,可以用于检测动物疫苗细胞培养物中支原体污染。  相似文献   

12.
DNA荧光染色和PCR技术在PRRSV冻干疫苗中支原体检测的应用   总被引:1,自引:0,他引:1  
为完善猪繁殖与呼吸综合征病毒(PRRSV)冻干疫苗生产各阶段的支原体检测体系,本研究分别应用DNA荧光染色法、PCR法和病毒分离培养法检测疫苗生产中的细胞、种毒、半成品、成品,分析方法的可行性和优势.结果显示,DNA荧光染色法检测PRRSV半成品抗原液的支原体污染能够于6d内得到结果,与其余半成品检验项目用时相近,适宜在生产中应用.而采用DNA荧光染色、PCR和病毒分离培养法对我们生产的10批PRRSV冻干疫苗成品的支原体检测结果显示,DNA荧光染色法与病毒分离培养法检测的结果一致,1批疫苗的PCR检测结果与培养法检测结果不符,即PCR法检测阳性,但DNA荧光染色和病毒分离培养法检测为阴性.由此证明,DNA荧光染色法和PCR法检测疫苗中支原体的结果可靠,而PCR法检出率更高.DNA荧光染色法和PCR法操作简便、快速、经济,可以作为疫苗生产质量的内控标准,提高支原体的检出率,保证疫苗质量.  相似文献   

13.
Tests for presence of mycoplasmas were conducted on 20 insemination bulls known as mycoplasma spreaders, with 5 sperm pellet batches of 20 pellets each being investigated for each animal. Mycoplasma contamination was positively recorded from 83 of the above 100 batches. Mycoplasma (M.) bovigenitalium, M. californicum. M. arginini, M. bovirhinis, and Acholeplasma laidlawii were typed by means of indirect immunofluorescent technique, which confirmed the presence also in the GDR of mycoplasma species described in the literature and detected in sperm samples. The solid culturing media used in the above tests, medium-B agar and cattle blood agar with staphylococcal nutrix, proved to be equally suitable for isolation of mycoplasma from sperm samples. Mycoplasma was positively identified in about one third of all pellets/batches tested. 3 pellets to one batch should be sufficient a random sample size from which to obtain information at least very close to real contamination.  相似文献   

14.
用清洗纯化法清除传代细胞系中支原体污染的方法探索   总被引:1,自引:0,他引:1  
为了解决引进细胞系的支原体污染和细胞传代生长不稳定问题,尝试对细胞培养条件和细胞群进行优选,并建立了清洗离心结合敏感抗生素法,达到了清除支原体和稳定细胞活力的目的.  相似文献   

15.
支原体PCR检测方法的建立及初步应用   总被引:1,自引:1,他引:0  
徐静 《中国兽药杂志》2013,47(11):17-21
经过对Genebank鸡滑液支原体、猪肺炎支原体、口腔支原体和猪鼻支原体16S rRNA序列比对,设计了一条通用引物,建立支原体PCR检测方法.该方法敏感、特异、经济、快速,在动物疫苗生产中检测细胞、半成品及成品支原体污染具有较大的应用价值.  相似文献   

16.
兽用疫苗中霉形体污染的套式PCR检测技术建立及初步应用   总被引:2,自引:0,他引:2  
根据GenBank公布的鸡毒霉形体、猪肺炎霉形体、猪滑液霉形体和絮状霉形体的16s rRNA基因序列,利用引物设计软件DNAStar和Primer5.0自行设计3对特异性引物,以市场上常见的兽用疫苗为检测对象进行试验,优化反应体系,建立了兽用疫苗霉形体污染检测的套式PCR方法。同时,本试验还对市场上的兽用疫苗进行随机抽查检测,其检出率分别为24.2%(23/95)、21.1%(20/95)和14.7%(14/95)。  相似文献   

17.
In order to establish a duplex PCR method for simultaneous detection of Mycoplasma ovipneumiae and Mycoplasma arginini, specific primers of Mycoplasma ovipneumiae and Mycoplasma arginini were designed, and evaluated its sensitivity and specificity after optimizing the reaction conditions of PCR.Then, a total of 40 nasal swabs were tested by duplex PCR.The assay could specifically amplify PCR fragments of 545 and 806 bp from Mycoplasma ovipneumiae and Mycoplasma arginini, respectively.While no PCR products were detected for other pathogens.The detection limits of the assay were determined to be 100 pg/μL for Mycoplasma ovipneumiae and 10 pg/μL for Mycoplasma arginini.The duplex PCR could detect Mycoplasma ovipneumiae and Mycoplasma arginini, and the coincidence rate could reach as high as 92.5% with enrichment culture about the 40 nasal swabs.The results suggested that the duplex PCR could be useful for clinical detection of Mycoplasma ovipneumiae and Mycoplasma arginini.  相似文献   

18.
为建立绵羊肺炎支原体和精氨酸支原体的双重PCR检测方法,本试验分别设计了绵羊肺炎支原体和精氨酸支原体的特异性引物,优化反应条件后对其特异性和敏感性进行评价,并对40份鼻拭子进行了检测。结果显示,该方法能同时扩增出绵羊肺炎支原体545 bp和精氨酸支原体806 bp的特异性片段,而对其他病原的DNA扩增均为阴性。该双重PCR方法对绵羊肺炎支原体和精氨酸支原体的最低检测限分别为100和10 pg/μL。40份鼻拭子检测结果显示,双重PCR检测方法与分离培养法符合率高达92.5%,均能鉴定出绵羊肺炎支原体和精氨酸支原体。结果表明,本研究建立的双重PCR方法可用于绵羊肺炎支原体和精氨酸支原体的临床快速诊断。  相似文献   

19.
The objective of this study was to determine the frequency of intraoperative contamination of the anterior chamber with viable microorganisms during cataract phacoemulsification and intraocular lens implantation, and to evaluate the relationship of contaminant microorganisms to patients' extraocular and nasal cavity floras. Also, the impact of various aspects of the patient history and phacoemulsification procedure on the incidence of positive postoperative anterior chamber cultures was investigated. Twenty-two eyes from 13 dogs presented for elective cataract phacoemulsification and intraocular lens implantation were studied. Preoperatively, microbiologic samples of the conjunctiva, eyelid margins, nares, and rostral nasal cavity were collected. Postoperatively, anterior chamber fluid was aspirated. Samples were submitted for aerobic/anaerobic bacteriologic culture and antimicrobial susceptibility, Mycoplasma culture, and fungal culture. Anterior chamber aspirates collected at the conclusion of surgery were culture positive for at least one organism in 22.7% of eyes. Three aerobic bacteria and three fungi were isolated from the anterior chamber aspirates. Two fungi and one bacterium isolated from the anterior chamber were typed identically, and the bacterium had a similar antibiogram to organisms recovered from the patient's conjunctiva and eyelid margin. No statistically significant difference in contamination frequency was found for the investigated patient and surgical variables. We conclude that intraoperative contamination of the anterior chamber with viable bacterial and fungal organisms is a common occurrence in canine patients undergoing cataract phacoemulsification and intraocular lens implantation, and the external ocular flora is a likely source of some of these contaminating microorganisms. This contamination is independent of the patient and surgical variables investigated.  相似文献   

20.
Between 1990 and 2000, more than 1600 mycoplasmas and the related acholeplasmas were identified from ruminant animals by the Mycoplasma Group at the Veterinary Laboratories Agency--Weybridge. Mycoplasma bovis was the most commonly identified pathogen, mostly from pneumonic calves but occasionally from cattle with mastitis and arthritis. Mycoplasma canis was first isolated in Britain in 1995 from pneumonic calves and the number of isolates increased to 18 per cent of the total mycoplasmas isolated from cattle in 1999. The ELISA for antibodies to M. bovis detected 1971 positive samples (22 per cent) among 8959 serum samples, mainly from pneumonic cattle. Other mycoplasmas identified included Mycoplasma dispar from the lungs of cattle with respiratory disease, and Mycoplasma bovigenitalium from the reproductive tract of cows with vulvovaginitis and infertility. Mycoplasma bovirhinis and Acholeplasma species were found commonly but are thought to be more opportunistic than pathogenic. In sheep and goats, the majority of Mycoplasma species isolated were identified as Mycoplasma ovipneumoniae from pneumonic sheep, Mycoplasma conjunctivae from sheep with keratoconjunctivitis, and the ubiquitous Mycoplasma arginini.  相似文献   

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