JP5704913B2 - リステリア属菌検出用簡易培地 - Google Patents
リステリア属菌検出用簡易培地 Download PDFInfo
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- JP5704913B2 JP5704913B2 JP2010284132A JP2010284132A JP5704913B2 JP 5704913 B2 JP5704913 B2 JP 5704913B2 JP 2010284132 A JP2010284132 A JP 2010284132A JP 2010284132 A JP2010284132 A JP 2010284132A JP 5704913 B2 JP5704913 B2 JP 5704913B2
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- listeria spp
- listeria
- glucopyranoside
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Landscapes
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Description
これらの培地を調製する際には、予め基礎となる培地成分を滅菌、保温し、これに抗生物質等からなる選択剤を添加後、分注、固化させ平板として使用することが一般的である。この場合、培地の調製及び検査のための検体の接種及び塗抹操作が非常に煩雑となっていた(非特許文献2〜4)。
(1)培地の作製
表1に記載の成分を本発明の培地組成物とした。この培地組成物のエタノール溶液1000mLに加え、懸濁液とした。このエタノール懸濁液を1mLずつ繊維質吸水性シート(コットン不織布、50φmm)を収納した容器(50φmm)に分注した後、これらが重ならないように、非開放空間内で一夜静置して乾燥した後、蓋をして本発明の簡易培地を作製した。本培地を乾燥剤と共にアルミ包材に密封包装した後、表面線量10〜20kGyのγ線照射を行って滅菌した。
トリプトソイ寒天(TSA)は1リットル使用量を1リットルの精製水に加え、121℃、15分間高圧蒸気滅菌し良く撹拌後、予め検体を接種したプラスチックシャーレ(90φmm)に15mLずつ分注した。その後、速やかに検体と培地を混合し、培地が固まるまで静置した。
供試菌株はトリプトソイブイヨンで24時間前培養したものを用い、これを滅菌生理食塩水で10倍段階希釈し、簡易培地及び空の滅菌プラスチックシャーレにはそれぞれ1mL、Oxford寒天培地及びALOA寒天培地平板には0.1mLをそれぞれ接種後、滅菌コンラージ棒を使用して塗抹を行った。
実際に本発明培地を使用し、リステリア属菌の検出を試みた結果、従来の培地を使用するよりも簡便に同等の結果を得ることができた(表2)。
また、リステリア属菌以外の菌株についてはそのほとんどが発育を抑えられる事を認めた(表2)。
(1)培地の作製
培地組成を表3に示した。
ALOA寒天培地に選択剤として使用されているポリミキシンBのみを選択剤として含むメッシュを有する繊維状吸水シートに担持させた構造を利用した簡易培地(実施例1と同様)をポリミキシンB含有量を変えてそれぞれ作製した。
供試菌株にはPseudomonas aeruginosa ATCC9027を使用し、トリプトソイブイヨンで24時間前培養したものを用いた。これを滅菌生理食塩水で10倍段階希釈し、簡易培地及び空の滅菌プラスチックシャーレにはそれぞれ1mL接種した。
ポリミキシンBを含有する簡易培地を作製した場合、通常寒天培地にて使用される濃度においては、その効力を発揮できず不活化していることを認め、寒天培地を同様の効果をもたらす為には少なくとも寒天培地に適用している濃度の10倍量が必要であることを認めた(表4)
従って、ポリミキシンBを選択剤に使用することは環境に対し、また安価に提供する点として適当でないことを認めた(表4)。
Claims (4)
- (a)キサンタンガム、(b)検出可能な遊離性基を有するβ−グルコシダーゼ基質、(c)硫酸リチウム、(d)コリスチン硫酸塩、(e)モクサラクタム及び(f)菌体栄養成分を含有する培地組成物を、繊維質吸水性シートに担持又は積層させたリステリア属菌検出用簡易培地。
- さらに、無機塩類及び/又は糖類を含有するものである請求項1記載のリステリア属菌検出用簡易培地。
- (b)検出可能な遊離性基を有するβ−グルコシダーゼ基質が、色原体化合物又は蛍光性化合物を遊離し得るβ−グルコピラノシド又はその塩である請求項1又は2記載のリステリア属菌検出用簡易培地。
- 請求項1〜3のいずれか1項記載の培地に、検体を接種して培養した後、当該培地上の検出可能なコロニーの有無を検出することを特徴とするリステリア属菌の検出法。
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