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猪胃样本中弓形杆菌属的检测

Detection of Arcobacter species in gastric samples from swine.

作者信息

Suarez D L, Wesley I V, Larson D J

机构信息

National Animal Disease Center, USDA/Agricultural Research Service, Ames, IA 50010, USA.

出版信息

Vet Microbiol. 1997 Oct 16;57(4):325-36. doi: 10.1016/s0378-1135(97)00107-7.

Abstract

Swine stomachs were surveyed for evidence of Arcobacter spp. and Helicobacter spp. infections associated with gastric ulceration. A nested PCR test targeted to the 16S rRNA was developed to detect many Arcobacter spp. and Helicobacter spp. An internal oligonucleotide probe was used for differentiation and confirmation of the PCR product. Tissue samples were obtained from the nonglandular and glandular regions of 86 swine stomachs. Evidence of infection with these microbes was detected in 51%, with 77% of the positive samples being identified as A. butzleri using a highly specific probe. Nonglandular stomach samples (44%) were more likely to be positive by PCR than samples from the glandular (23%) region. Gross lesions of any stage of gastric ulceration, ranging from parakeratosis, erosions and ulceration, were observed in 24% of stomachs examined. Of 21 samples with lesions, 52% were positive by the broadly reactive PCR assay for Arcobacter spp. and Helicobacter spp. The majority of PCR-positive samples (75%) had no gross lesions. When a single step PCR assay that was more specific for Arcobacter spp. was used on the nonglandular stomach samples, 10.4% of the 86 samples were positive. Arcobacter spp. were cultured from four of the sample stomachs. Partial sequencing of the 16S rRNA gene identified the isolates as A. butzleri (n = 2), A. cryaerophilus, (n = 1), and a mixed culture of A. butzleri and another Arcobacter spp. (n = 1). A single step PCR assay targeted to the urease gene and culturing methods were used to screen for H. pylori or other closely related urease positive bacteria, but none were found.

摘要

对猪胃进行了调查,以寻找与胃溃疡相关的弓形杆菌属和螺杆菌属感染的证据。开发了一种针对16S rRNA的巢式PCR检测方法,以检测多种弓形杆菌属和螺杆菌属。使用内部寡核苷酸探针来区分和确认PCR产物。从86个猪胃的非腺区和腺区获取组织样本。在51%的样本中检测到了这些微生物的感染证据,其中77%的阳性样本使用高度特异性探针鉴定为布氏弓形杆菌。非腺胃样本(44%)通过PCR检测呈阳性的可能性高于腺区样本(23%)。在所检查的胃中,24%观察到了任何阶段的胃溃疡肉眼病变,范围从角化不全、糜烂到溃疡。在21个有病变的样本中,52%通过针对弓形杆菌属和螺杆菌属的广泛反应性PCR检测呈阳性。大多数PCR阳性样本(75%)没有肉眼病变。当对非腺胃样本使用对弓形杆菌属更具特异性的单步PCR检测时,86个样本中有10.4%呈阳性。从4个样本胃中培养出了弓形杆菌属。16S rRNA基因的部分测序将分离株鉴定为布氏弓形杆菌(n = 2)、嗜低温弓形杆菌(n = 1)以及布氏弓形杆菌与另一种弓形杆菌属的混合培养物(n = 1)。使用针对脲酶基因的单步PCR检测和培养方法来筛查幽门螺杆菌或其他密切相关的脲酶阳性细菌,但未发现。

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