A sensitive and specific PCR method to detect Helicobacter felis in a conventional mouse model.

L Kong; J G Smith; D Bramhill; G K Abruzzo; C Bonfiglio; C Cioffe

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A sensitive and specific PCR method to detect Helicobacter felis in a conventional mouse model.

机译：一种灵敏且特异性的PCR方法，可检测常规小鼠模型中的猫幽门螺杆菌。

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Although many detection methods have been used to determine Helicobacter colonization in small animal models, the sensitivity and specificity of these detection methods are limited. To improve the Helicobacter felis conventional mouse model for accurate evaluation of therapeutic regimens, we developed a PCR for detection of, and a competitive PCR for quantitation of, H. felis in viral antibody-free (VAF) mice. The PCR was based on the H. felis 16S rRNA gene. An internal control DNA was used for competitive quantitation of the PCR. VAF conventional Swiss-Webster mice were infected with an H. felis culture by oral gavage. At various times after H. felis challenge and therapy, stomach mucosa was collected and evaluated by PCR. PCR detected approximately 50 to 100 H. felis cells per mouse stomach and showed no cross-reaction with other bacteria commonly found in mouse stomachs. Colonization of H. felis in the mouse stomach was confirmed by culture isolation from germfree mice and histological examination of VAF mice. Response to therapy in this H. felis model correlated well with results seen in human clinical trials with H. pylori. A model utilizing PCR detection which may be useful for discovering new antibiotics and/or vaccines against Helicobacter ulcer disease has been developed.

机译：尽管在小动物模型中已使用许多检测方法来确定幽门螺杆菌定植，但是这些检测方法的敏感性和特异性受到限制。为了改进用于准确评估治疗方案的猫幽门螺杆菌常规小鼠模型，我们开发了一种用于检测无病毒抗体（VAF）小鼠中猫幽门螺杆菌的PCR和一种竞争性PCR定量方法。 PCR基于H. felis 16S rRNA基因。内部对照DNA用于PCR的竞争定量。通过口腔管饲法将VAF常规的Swiss-Webster小鼠感染了H. felis培养物。在H. felis攻击和治疗后的不同时间，收集胃粘膜并通过PCR进行评估。 PCR检测到每个小鼠胃中大约有50至100个H. felis细胞，并且与小鼠胃中常见的其他细菌没有交叉反应。通过从无菌小鼠中分离培养物和对VAF小鼠进行组织学检查，证实了H. felis在小鼠胃中的定殖。在此H. felis模型中对治疗的反应与幽门螺杆菌在人体临床试验中看到的结果密切相关。已经开发出利用PCR检测的模型，该模型可用于发现针对幽门螺杆菌疾病的新抗生素和/或疫苗。

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《Clinical and diagnostic laboratory immunology》 |1996年第1期|共6页
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L Kong; J G Smith; D Bramhill; G K Abruzzo; C Bonfiglio; C Cioffe;
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A sensitive and specific PCR method to detect Helicobacter felis in a conventional mouse model.

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