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通过在滤纸条点试验中使用2'-(4-甲基伞形酮基)-α-D-N-乙酰神经氨酸检测放线菌属中的唾液酸酶(神经氨酸酶)活性。

Detection of sialidase (neuraminidase) activity in Actinomyces species by using 2'-(4-methylumbelliferyl)alpha-D-N-acetylneuraminic acid in a filter paper spot test.

作者信息

Moncla B J, Braham P

机构信息

Department of Oral Biology, University of Washington, Seattle 98195.

出版信息

J Clin Microbiol. 1989 Jan;27(1):182-4. doi: 10.1128/jcm.27.1.182-184.1989.

Abstract

A rapid method for the detection of acetylneuraminyl hydrolase, EC 3.2.1.18 (sialidase or neuraminidase), was developed by using 2'-(4-methylumbelliferyl)alpha-D-N-acetylneuraminic acid as substrate in a filter paper spot test. The method was compared to conventional assays that use 2'-(4-methylumbelliferyl)alpha-D-N-acetylneuraminic acid and bovine submaxillary mucin and was found to be in excellent agreement. Organisms with greater than 10 U of enzyme activity (in nanomoles per minute per milligram of cell protein) gave positive reactions, while those with 2.7 to 9.0 U gave only weak reactions. Isolates with less than 2.7 U of activity were detected upon prolonged incubation. Sialidase activity was detected in 79% of 71 clinical isolates representing five species of Actinomyces. The percentage of sialidase-producing isolates of each species varied considerably: Actinomyces israelii, 63%; A. meyeri, 73%; A. naeslundii, 85%; A. odontolyticus, 73%; and A. viscosus, 100%.

摘要

通过在滤纸斑点试验中使用2'-(4-甲基伞形酮基)-α-D-N-乙酰神经氨酸作为底物,开发了一种快速检测乙酰神经氨酸水解酶(EC 3.2.1.18,唾液酸酶或神经氨酸酶)的方法。将该方法与使用2'-(4-甲基伞形酮基)-α-D-N-乙酰神经氨酸和牛颌下粘蛋白的传统检测方法进行比较,发现两者具有极佳的一致性。酶活性大于10 U(每毫克细胞蛋白每分钟纳米摩尔数)的微生物产生阳性反应,而酶活性为2.7至9.0 U的微生物仅产生微弱反应。活性低于2.7 U的分离株在延长孵育后被检测到。在代表五种放线菌的71株临床分离株中,79%检测到唾液酸酶活性。每种产生唾液酸酶的分离株的百分比差异很大:以色列放线菌为63%;迈耶放线菌为73%;内氏放线菌为85%;溶齿放线菌为73%;粘性放线菌为100%。

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本文引用的文献

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Adv Carbohydr Chem Biochem. 1982;40:131-234. doi: 10.1016/s0065-2318(08)60109-2.
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Curr Top Microbiol Immunol. 1972;59:35-74. doi: 10.1007/978-3-642-65444-2_2.

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