通过热休克诱导的mRNA扩增检测活的贾第虫囊肿
Detection of viable Giardia cysts by amplification of heat shock-induced mRNA.
作者信息
Abbaszadegan M, Huber M S, Gerba C P, Pepper I L
机构信息
Quality Control & Research Laboratory, American Water Works Service Company, Inc., Belleville, Illinois 62220, USA.
出版信息
Appl Environ Microbiol. 1997 Jan;63(1):324-8. doi: 10.1128/aem.63.1.324-328.1997.
Abstract
Primers obtained from gene sequences coding for heat shock proteins (HSP) were used to specifically detect enteric protozoans of the genus Giardia. The HSP primers amplified Giardia DNA or the corresponding RNA sequences obtained from lysed cysts and gave a 163-bp product. Since the presence of the product did not indicate whether the cysts were viable, these amplifications are a presence/absence test only. In contrast, amplification of heat shock-induced mRNA utilizing the same HSP primers was indicative of viable Giardia cysts. The limit of sensitivity of the presence/absence test was 1 cyst, whereas for the viability test it was 10 cysts. Thus, viable Giardia cysts can be rapidly and specifically detected with great sensitivity through the use of PCR amplifications.
摘要
从编码热休克蛋白（HSP）的基因序列中获得的引物被用于特异性检测贾第虫属的肠道原生动物。HSP引物扩增了从裂解囊肿中获得的贾第虫DNA或相应的RNA序列，并产生了一个163bp的产物。由于产物的存在并不表明囊肿是否存活，这些扩增仅为存在/不存在检测。相比之下，利用相同的HSP引物扩增热休克诱导的mRNA则表明贾第虫囊肿是存活的。存在/不存在检测的灵敏度极限为1个囊肿，而活力检测的灵敏度极限为10个囊肿。因此，通过使用PCR扩增，可以快速、特异性且高度灵敏地检测存活的贾第虫囊肿。
相似文献
1
Detection of viable Giardia cysts by amplification of heat shock-induced mRNA.通过热休克诱导的mRNA扩增检测活的贾第虫囊肿
Appl Environ Microbiol. 1997 Jan;63(1):324-8. doi: 10.1128/aem.63.1.324-328.1997.
2
Comparison of primers and optimization of PCR conditions for detection of Cryptosporidium parvum and Giardia lamblia in water.用于检测水中微小隐孢子虫和蓝氏贾第鞭毛虫的引物比较及聚合酶链反应条件优化
Appl Environ Microbiol. 1997 Jan;63(1):106-14. doi: 10.1128/aem.63.1.106-114.1997.
3
Giardia duodenalis: improved detection of viable cysts by reverse transcription-PCR of heat shock-inducible hsp70 gene.十二指肠贾第虫：通过热休克诱导型hsp70基因的逆转录聚合酶链反应改进对活囊蚴的检测
Exp Parasitol. 2009 Dec;123(4):377-80. doi: 10.1016/j.exppara.2009.08.011. Epub 2009 Aug 22.
4
[New methods for the diagnosis of Cryptosporidium and Giardia].[隐孢子虫和贾第虫的诊断新方法]
Parassitologia. 2004 Jun;46(1-2):151-5.
5
Detection of Giardia cysts by using the polymerase chain reaction and distinguishing live from dead cysts.利用聚合酶链反应检测贾第虫包囊并区分活包囊与死包囊。
Appl Environ Microbiol. 1991 Dec;57(12):3456-61. doi: 10.1128/aem.57.12.3456-3461.1991.
6
ECC-RT-PCR: a new method to determine the viability and infectivity of Giardia cysts.ECC-RT-PCR：一种用于确定贾第虫包囊活力和感染力的新方法。
Int J Infect Dis. 2012 May;16(5):e350-3. doi: 10.1016/j.ijid.2012.01.004. Epub 2012 Mar 3.
7
Immunomagnetic separation significantly improves the sensitivity of polymerase chain reaction in detecting Giardia duodenalis and Cryptosporidium spp. in dairy cattle.免疫磁珠分离法显著提高了聚合酶链反应检测奶牛十二指肠贾第虫和隐孢子虫的灵敏度。
J Vet Diagn Invest. 2011 Mar;23(2):260-7. doi: 10.1177/104063871102300210.
8
Improved specificity for Giardia lamblia cyst quantification in wastewater by development of a real-time PCR method.通过开发实时聚合酶链反应方法提高废水中华支睾吸虫囊肿定量的特异性。
J Microbiol Methods. 2004 Apr;57(1):41-53. doi: 10.1016/j.mimet.2003.11.016.
9
Sequence analysis of the beta-giardin gene and development of a polymerase chain reaction-restriction fragment length polymorphism assay to genotype Giardia duodenalis cysts from human faecal samples.β-贾第虫基因的序列分析及聚合酶链反应-限制性片段长度多态性分析方法的建立，用于对人粪便样本中的十二指肠贾第虫囊肿进行基因分型。
Int J Parasitol. 2002 Jul;32(8):1023-30. doi: 10.1016/s0020-7519(02)00068-1.
10
Simplified protocol for DNA extraction and amplification of 2 molecular markers to detect and type Giardia duodenalis.检测和分型贾第虫的 2 种分子标记物的 DNA 提取和扩增简化方案。
Diagn Microbiol Infect Dis. 2014 Jan;78(1):53-8. doi: 10.1016/j.diagmicrobio.2013.09.008. Epub 2013 Sep 28.
引用本文的文献
1
Detection of spp. and spp. in Environmental Water Samples: A Journey into the Past and New Perspectives.环境水样中 spp. 和 spp. 的检测：回顾与新视角
Microorganisms. 2022 Jun 7;10(6):1175. doi: 10.3390/microorganisms10061175.
2
Assessing viability and infectivity of foodborne and waterborne stages (cysts/oocysts) of Giardia duodenalis, Cryptosporidium spp., and Toxoplasma gondii: a review of methods.评估十二指肠贾第虫、隐孢子虫属和刚地弓形虫的食源性和水源性阶段（包囊/卵囊）的活力和感染性：方法综述
Parasite. 2018;25:14. doi: 10.1051/parasite/2018009. Epub 2018 Mar 19.
3
Sensitivity of nested PCR in the detection of low numbers of Giardia lamblia cysts.巢式聚合酶链反应检测少量蓝氏贾第鞭毛虫包囊的灵敏度
Appl Environ Microbiol. 2007 Sep;73(18):5949-50. doi: 10.1128/AEM.00668-07. Epub 2007 Jul 20.
4
Specific detection of DNA and RNA targets using a novel isothermal nucleic acid amplification assay based on the formation of a three-way junction structure.基于三链交联结构形成的新型等温核酸扩增分析法对DNA和RNA靶标的特异性检测。
Nucleic Acids Res. 2001 Jun 1;29(11):E54-4. doi: 10.1093/nar/29.11.e54.
5
Viability of Giardia intestinalis cysts and viability and sporulation state of Cyclospora cayetanensis oocysts determined by electrorotation.通过电旋转法测定小肠贾第虫包囊的活力以及卡耶塔环孢子虫卵囊的活力和孢子化状态。
Appl Environ Microbiol. 2001 Feb;67(2):586-90. doi: 10.1128/AEM.67.2.586-590.2001.
6
Development and evaluation of a molecular viability assay for Pneumocystis carinii.卡氏肺孢子虫分子活力测定法的开发与评估
J Clin Microbiol. 2000 May;38(5):1947-52. doi: 10.1128/JCM.38.5.1947-1952.2000.
7
Sensitive and rapid detection of viable Giardia cysts and Cryptosporidium parvum oocysts in large-volume water samples with wound fiberglass cartridge filters and reverse transcription-PCR.使用伤口玻璃纤维筒式过滤器和逆转录聚合酶链反应灵敏快速地检测大量水样中的活贾第虫包囊和微小隐孢子虫卵囊。
Appl Environ Microbiol. 1998 May;64(5):1743-9. doi: 10.1128/AEM.64.5.1743-1749.1998.
本文引用的文献
1
The experimental transmission of human intestinal protozoan parasites. II. Giardia lamblia cysts given in capsules.人体肠道原生动物寄生虫的实验性传播。II. 以胶囊形式给予的蓝氏贾第鞭毛虫包囊。
Am J Hyg. 1954 Mar;59(2):209-20. doi: 10.1093/oxfordjournals.aje.a119634.
2
Municipal waterborne giardiasis: an epidemilogic investigation. Beavers implicated as a possible reservoir.城市水源性贾第虫病：一项流行病学调查。海狸被认为可能是储存宿主。
Ann Intern Med. 1980 Feb;92(2 Pt 1):165-70. doi: 10.7326/0003-4819-92-2-165.
3
The heat shock response is self-regulated at both the transcriptional and posttranscriptional levels.热休克反应在转录和转录后水平上都是自我调节的。
Cell. 1982 Dec;31(3 Pt 2):593-603. doi: 10.1016/0092-8674(82)90315-4.
4
Improved in vitro excystation procedure for Giardia lamblia cysts.改进的蓝氏贾第鞭毛虫囊肿体外脱囊程序。
J Clin Microbiol. 1981 Dec;14(6):709-10. doi: 10.1128/jcm.14.6.709-710.1981.
5
Giardia lamblia infections in Mongolian gerbils: an animal model.蒙古沙鼠贾第虫感染：一种动物模型。
J Infect Dis. 1983 Feb;147(2):222-6. doi: 10.1093/infdis/147.2.222.
6
Translational efficiency of heat-induced messages in Drosophila melanogaster cells.黑腹果蝇细胞中热诱导信息的翻译效率。
J Mol Biol. 1980 Feb 25;137(2):151-8. doi: 10.1016/0022-2836(80)90322-8.
7
Comparison of animal infectivity and excystation as measures of Giardia muris cyst inactivation by chlorine.比较动物感染性和脱囊作用作为衡量氯对鼠贾第虫囊肿灭活效果的指标。
Appl Environ Microbiol. 1985 Oct;50(4):1115-7. doi: 10.1128/aem.50.4.1115-1117.1985.
8
Use of immunofluorescence and phase-contrast microscopy for detection and identification of Giardia cysts in water samples.利用免疫荧光和相差显微镜检测与鉴定水样中的贾第虫包囊。
Appl Environ Microbiol. 1985 Dec;50(6):1434-8. doi: 10.1128/aem.50.6.1434-1438.1985.
9
A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation.GM-CSF信使核糖核酸3'非翻译区的一段保守AU序列介导选择性信使核糖核酸降解。
Cell. 1986 Aug 29;46(5):659-67. doi: 10.1016/0092-8674(86)90341-7.
10
Epidemic giardiasis caused by a contaminated public water supply.由受污染的公共供水系统引起的流行性贾第虫病。
Am J Public Health. 1988 Feb;78(2):139-43. doi: 10.2105/ajph.78.2.139.
Zotero 插件