共查询到20条相似文献，搜索用时 109 毫秒
1.
根椐Gen Bank公布的兔多杀性巴氏杆菌16S r RNA和支气管败血波氏杆菌fim N基因序列,设计两对引物,在建立两种细菌单项PCR检测方法的基础上,优化双重PCR反应条件,建立了两种细菌的双重PCR检测方法,用这两对引物对同一样品中的兔多杀性巴氏杆菌、支气管败血波氏杆菌核酸为模板进行双重PCR扩增,结果可同时扩增兔多杀性巴氏杆菌和支气管败血波氏杆菌的258bp和449bp的特异性片段,而对其他4种兔病原菌的扩增结果均为阴性。敏感性测定结果表明,对兔多杀性巴氏杆菌和支气管败血波氏杆菌的最低核酸检出限分别均为1pg。通过对78份临床病料检测,将建立的双重PCR技术和单项PCR方法进行对比验证,结果显示,两者的总符合率为100%。表明建立的双重PCR检测方法,具有特异、快速、准确的特点,可用于对这兔多杀性巴氏杆菌和支气管败血波氏杆菌的同时检测和鉴别诊断。 相似文献
2.
3.
《中国预防兽医学报》2018,(10)
为建立支气管败血波氏杆菌SYBR GreenⅠ荧光定量PCR检测方法,本研究根据波氏菌属毒力基因DNT设计一对特异性引物,通过优化,确定最佳反应条件,建立了支气管败血波氏杆菌荧光定量PCR检测方法。并对其进行敏感性、特异性、重复性试验以及临床样品的检测。以支气管败血波氏杆菌标准质粒为模板,建立标准曲线结果显示:标准品浓度在1.13×10~8拷贝/μL～1.13×10~2拷贝/μL范围内具有良好的线性关系,相关数R~2=0.997;该方法的灵敏度可达1.13×10~1拷贝/μL。建立的荧光定量PCR方法与副猪嗜血杆菌、猪链球菌、金黄色葡萄球菌等12种细菌和伪狂犬病毒、猪瘟病毒等4种病毒无交叉反应,具有良好的特异性;批内变异系数(CV)均小于2%,批间变异系数均小于4%,具有良好的稳定性。对临床样品的检测结果显示该方法检测的阳性率为57.7%,与普通PCR的符合率为93.24%。本研究建立的方法可以用作支气管败血波氏杆菌感染的早期诊断、快速检测与定量分析。 相似文献
4.
《中国预防兽医学报》2015,(9)
为建立一种快速检测蜜蜂黑蜂王台病毒(BQCV)的环介导等温扩增方法(LAMP),本研究根据Gen Bank中登录的BQCV-JL1株基因序列(KP119603.1)设计了多套LAMP引物,通过引物筛选,反应体系和反应条件优化,建立了可视化LAMP方法,并且评价了该方法的特异性、敏感性、重复性和稳定性。结果显示,建立的LAMP方法在63℃下可对BQCV核酸进行高效扩增,可视化判定结果与real-time Turbidimeter仪检测结果一致;该方法具有较强的特异性和较高的敏感性(最低检出限量为4.0×102拷贝/μL),为普通PCR的100倍。临床样品检测结果表明LAMP法检出率高于常规PCR。本研究建立的可视化LAMP方法操作简便,适用于BQCV的快速检测。 相似文献
5.
为建立一种快速准确检测兔多杀性巴氏杆菌(P.multocida)的PCR方法,本研究以P.multocida的高度保守的16S rRNA为靶基因,参考已公布的P.multocida的16S rRNA基因设计1对特异性引物,优化PCR反应条件,建立了P.multocida PCR快速检测方法。该PCR方法的敏感性达到60 cfu/mL,采用该PCR方法扩增P.multocida标准株和分离株均能扩增出643 bp的目的片段,扩增兔大肠杆菌、支气管败血波氏杆菌结果为阴性,证明本实验所建立的P.multocida PCR检测方法快速、敏感、特异、可靠,可用于P.multocida的快速鉴定与诊断。同时用建立的PCR方法对临床疑似病兔的脏器分离菌进行扩增,可扩增出目的条带,与细菌的分离结果相一致。 相似文献
6.
参考GenBank中兔巴氏杆菌16SrRNA和波氏杆菌的fim2的基因序列,应用Premier 5.0软件在二者高度保守区设计了2对引物,建立了适合巴氏杆菌和波氏杆菌快速检测的多重PCR检测方法。以该方法对巴氏杆菌和波氏杆菌参考菌株进行PCR扩增,分别能从各自的基因组中扩增出与试验设计相符的644bp和425bp的特异性DNA片段。将扩增所得的DNA片段进行克隆测序,测序结果表明分别为巴氏杆菌16SrRNA和波氏杆菌fim2基因序列。该方法对波氏杆菌的检测下限为4×102 CFU,对巴氏杆菌的检测下限为6×101 CFU。对兔源性沙门菌、葡萄球菌、大肠杆菌、魏氏梭菌扩增结果为阴性。表明所建立的RT-PCR检测技术具有特异、快速和敏感的特点,可用于鉴别诊断兔巴氏杆菌和波氏杆菌以及2者混合感染。 相似文献
7.
动物饲料中沙门氏菌环介导等温扩增(LAMP)快速诊断方法的建立 总被引：1，自引：0，他引：1
根据GenBank公布的沙门氏菌高度保守的fimY蛋白基因序列,利用Primer explorer V4软件设计4条针对沙门氏菌fimY基因的特异性引物,通过对Mg2+浓度、dNTP浓度、反应温度、反应时间、特异性、敏感性、重复性和稳定系及符合率等方面,优化LAMP各种反应条件,建立针对沙门氏菌LAMP快速诊断方法。结果表明,该方法具有较强的特异性;比PCR检测方法敏感性高100倍,对沙门氏菌的最低检出量为6.2 CFU/mL;对24份样品的3次检测结果与国标法完全一致。证明建立的针对fimY蛋白基因的LAMP检测方法操作简便,特异性强,敏感性高,适用于现场检测。 相似文献
8.
联合检测兔巴氏杆菌和波氏杆菌病血清抗体的Dot-ELISA方法研究 总被引：2，自引：0，他引：2
兔多杀性巴氏杆菌病(Pm)和兔支气管败血波氏杆菌病(Bb)是家兔的两种主要细菌性传染病。兔波氏杆菌病可引起家兔的慢性鼻炎,兔巴氏杆菌病也可引起包括呼吸道在内的多种疾病。试验旨在建立一种简便、快速、经济、准确的联合检测Pm和Bb抗体水平的方法,可用于这两种疾病的免疫监测及流行病学调查。1材料1.1菌种兔巴氏杆菌Pm90株、兔支气管败血波氏杆菌Bb82株,为山东省农业科学院畜牧兽医研究所分离保存[1]。1.2血清大肠杆菌多因子血清、沙门菌多因子血清,购自中国兽药监察所;魏氏梭菌、兔瘟、Pm和Bb阳性血清,山东省农业科学院畜牧兽医研究所… 相似文献
9.
本试验以超声破碎和高速离心方法提取禽波氏杆菌外膜蛋白,作为抗原免疫家兔,制备并纯化兔抗禽波氏杆菌外膜蛋白IgG,建立了能进行禽波氏杆菌抗原定位和检测的间接免疫酶组化法,并对试验条件进行了优化。对禽波氏杆菌人工感染海兰褐雏鸡的检测结果表明,该法能特异性地对气管、肺、肝、心、脾、肾中的禽波氏杆菌进行抗原定位,同细菌分离鉴定的结果符合率为100％。同时对感染鸡胚肝脏和心脏中禽波氏杆菌的检测结果均为阳性。该法具有高度敏感性和特异性特点,可用于禽波氏杆菌临床和试验感染条件下的诊断和定位检测。 相似文献
10.
兔支气管败血波氏杆菌PCR检测方法的建立及应用 总被引：2，自引：0，他引：2
根据支气管败血波氏杆菌菌毛蛋白基因(fimN)序列设计了一对引物,扩增出大小为648bp的目的基因片段,建立了快速检测兔支气管败血波氏杆菌的PCR方法。特异性和敏感性试验表明,该方法对大肠埃希氏菌、金黄色葡萄球菌和巴氏杆菌均无交叉性反应。用该PCR法检测了从杨凌、咸阳采集的50份表现临床症状的兔呼吸道分泌物检出支气管败血波氏杆菌阳性35例,阳性率为70%。同时与传统的细菌检查法、微量凝集法进行了比较,结果显示,该PCR法的检出率是细菌检查法的3.89倍,是微量凝集法的1.94倍。 相似文献
11.
An 18‐year‐old stallion was presented for castration because of insidious, bilateral scrotal enlargement of one year's duration. The left testis was firm, while the right was soft and lobulated; both were larger than normal. Palpation of the scrotum and its contents did not cause the horse to show signs of discomfort. Ultrasonography of scrotal contents revealed abnormal, heterogeneous tissue with ill defined regions of hyper‐ and hypoechogenity throughout both testes. Several hours after admission the horse developed severe signs of colic. On the basis of anamnesis, clinical findings, and results of ultrasonography and transrectal palpation bilateral testicular neoplasia and incarcerated inguinal hernia were tentatively diagnosed. Because the horse's owner did not consent to surgical treatment, the stallion was subjected to euthanasia. Histological and immunohistochemical examination of the testicular tissue collected post mortem revealed a seminoma in the left testis and a leiomyoma in the right testis. The post mortem examination also revealed incarceration of the small intestine in addition to the testicular tumours. To our knowledge, this is the first report of the simultaneous presence of bilateral, yet different testicular tumour types in a stallion. 相似文献
12.
13.
Laluha P Grest P Eichenberger S Reusch CE Gerber B 《Schweizer Archiv für Tierheilkunde》2006,148(6):303-307
A five-year-old, neutered male, Bernese Mountain Dog was referred because of a two-week history of hematuria and a one-week history of polydipsia and polyuria. Palpation of the dorsomedial mesogastrium revealed a mass on the left side, which was seen via radiography and ultrasonography to be attached to the left kidney. The left kidney and the attached mass were excised surgically, and the dog had an uneventful recovery. Histological examination of the mass revealed a dense proliferation of non-polymorphic spindle cells, which stained positive for smooth muscle actin via immunohistochemistry. Leiomyoma of the kidney was diagnosed. Postoperatively, there was mild azotaemia, which did not affect the general condition of the dog and was still present 15 months after surgery. Leiomyoma of the kidney is a benign tumor, and hematuria may be the only clinical sign. 相似文献
14.
15.
16.
M FLANAGAN 《Australian veterinary journal》1992,69(5):112-113
17.
18.
19.
Surgical resection of a dysgerminoma in a mare 总被引：1，自引：1，他引：0
A mare was referred for further evaluation of a mass found in the left caudal abdomen during a routine postpartum reproductive palpation. The mare was clinically normal with no history of health problems. Ultrasonographic examination of the mass confirmed its presence, but the origin of the mass could not be accurately determined. Routine haematology and serum biochemistry results were within normal limits. The mare was initially treated conservatively with antibiotics, but the mass continued to increase in size, so it was surgically excised. The mass involved the left ovary. The mare showed transient abdominal pain after surgery, but developed no other complications and was in foal 7 months later. On histology, the mass was diagnosed as a dysgerminoma, a rare ovarian tumour of germ cell origin. 相似文献
20.
Brennan SF Connery N Tobin E Mooney CT Jones BR 《The Journal of small animal practice》2004,45(6):304-306
A six-year-old, female Tibetan terrier was referred for investigation of a non-healing wound on the left caudal thorax. A subcutaneous swelling had initially developed on the chest wall, followed by a draining tract from which seropurulent fluid drained for two months. There had been no response to antibiotic treatment. Following radiographic and ultrasonographic examinations, a bone sequestrum from a fractured rib or a foreign body was suspected. Surgical exploration of the wound identified a sinus tract and a wooden foreign body (an ice-lolly stick) was located in subcutaneous tissues. Partial wound dehiscence of the surgical site occurred postoperatively, but healed after 10 days. One month later, fluid began to discharge from the area again. Further surgical exploration confirmed a gastrocutaneous fistula. Dissection of the fistula and surgical closure of the stomach, body wall and skin led to resolution of all signs. 相似文献