PL423812A1 - Method for producing colorimetric biosensor for detection of bacteria of Escherichia coli species and the colorimetric biosensor for detection of bacteria of Escherichia coli species - Google Patents
Method for producing colorimetric biosensor for detection of bacteria of Escherichia coli species and the colorimetric biosensor for detection of bacteria of Escherichia coli speciesInfo
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- PL423812A1 PL423812A1 PL423812A PL42381217A PL423812A1 PL 423812 A1 PL423812 A1 PL 423812A1 PL 423812 A PL423812 A PL 423812A PL 42381217 A PL42381217 A PL 42381217A PL 423812 A1 PL423812 A1 PL 423812A1
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- biosensor
- buffer
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- detection
- escherichia coli
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Abstract
Przedmiotem zgłoszenia jest sposób wytwarzania biosensora kolorymetrycznego do detekcji bakterii z gatunku Escherichia coli, który charakteryzuje się tym, że przeprowadzając, znanymi metodami, precypitację soli żelaza w środowisku alkalicznym otrzymuje się ferrofluid modyfikowany polietylenoiminą, stanowiący matrycę biosensora, którą płucze się co najmniej trzy razy wodą dejonizowaną stabilizując pH na poziome równym 7, po czym dodaje się bakteriofagi T4 w ilości 109 na 1 mg matrycy i inkubuje się co najmniej 12 godzin w temperaturze od 5 - 10°C z wytrząsaniem. Następnie magnetycznie odseparowuje się cząstki ferromagnetyczne niosące bakteriofaga T4, usuwa supernatant, płucze co najmniej pięciokrotnie buforem SM i dodaje się roztwór nasycony albuminy bydlęcej BSA w ilości 1 mg albuminy na 1 mL cząstek ferromagnetycznych z bakteriofagami T4 i tak otrzymany biosensor (1) inkubuje się co najmniej 5 godzin w temperaturze pokojowej i płucze się buforem SM co najmniej 3 razy. Biosensor (1) w ilości co najmniej 100 mg umieszcza się w przeźroczystym naczyniu szklanym (3), w buforze (2) o składzie: czerwień fenolowa 0,5, mM, laktoza 100 mM. Przedmiotem zgłoszenia jest zestaw do detekcji bakterii z gatunku Escherichia coli, który charakteryzuje się tym, że składa się z biosensora (1) kolorymetrycznego w postaci matrycy z ferrofluidu modyfikowanego polietylenoiminą z zimmobilizowanymi na niej bakteriofagami T4 w ilości 109 wirionów/100 mg matrycy, który to biosensor (1) umieszczony jest w przeźroczystym naczyniu szklanym (3), w buforze (2), o składzie: czerwień fenolowa 0,5 mM, laktoza 100 mM oraz z magnesu kołowego (4) umożliwiającego odseparowanie złoża w trakcie procedury detekcji. Ilość biosensora (1) w naczyniu szklanym wynosi co najmniej 100 mg, zaś ilość buforu (2) co najmniej 0,5 mL. Zestaw może zawierać dodatkowo drugie naczynie szklane (3a), w którym znajduje się próba negatywna.The subject of the application is a method for producing a colorimetric biosensor for the detection of Escherichia coli bacteria, which is characterized by the fact that by known methods, precipitation of iron salts in an alkaline environment, a ferrofluid modified with polyethyleneimine is obtained, which is a biosensor matrix which is washed at least three times with water deionized stabilizing the pH at 7, then T4 bacteriophages are added in an amount of 109 per 1 mg matrix and incubated for at least 12 hours at a temperature of 5 - 10 ° C with shaking. Then the ferromagnetic particles carrying the T4 bacteriophage are magnetically separated, the supernatant is removed, the buffer is washed at least five times with SM buffer and a saturated solution of bovine albumin BSA is added in an amount of 1 mg albumin per 1 mL of ferromagnetic particles with T4 bacteriophages and the biosensor (1) is incubated at least 5 hours at room temperature and washed with SM buffer at least 3 times. The biosensor (1) in an amount of at least 100 mg is placed in a transparent glass dish (3), in buffer (2) composed of phenol red 0.5, mM, lactose 100 mM. The subject of the application is the Escherichia coli bacterial detection kit, which is characterized by the fact that it consists of a (1) colorimetric biosensor in the form of a polyethyleneimine modified ferrofluid matrix with T4 bacteriophages immobilized on it in the amount of 109 virions / 100 mg matrix, which the biosensor (1) is placed in a transparent glass vessel (3), in a buffer (2), composed of: phenol red 0.5 mM, lactose 100 mM and a circular magnet (4) enabling separation of the bed during the detection procedure. The amount of biosensor (1) in the glass dish is at least 100 mg, and the amount of buffer (2) is at least 0.5 mL. The kit may additionally contain a second glass container (3a) in which the negative test is contained.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PL423812A PL236550B1 (en) | 2017-12-11 | 2017-12-11 | Method for producing colorimetric biosensor for detection of bacteria of Escherichia coli species and the colorimetric biosensor for detection of bacteria of Escherichia coli species |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PL423812A PL236550B1 (en) | 2017-12-11 | 2017-12-11 | Method for producing colorimetric biosensor for detection of bacteria of Escherichia coli species and the colorimetric biosensor for detection of bacteria of Escherichia coli species |
Publications (2)
Publication Number | Publication Date |
---|---|
PL423812A1 true PL423812A1 (en) | 2019-06-17 |
PL236550B1 PL236550B1 (en) | 2021-01-25 |
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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PL423812A PL236550B1 (en) | 2017-12-11 | 2017-12-11 | Method for producing colorimetric biosensor for detection of bacteria of Escherichia coli species and the colorimetric biosensor for detection of bacteria of Escherichia coli species |
Country Status (1)
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PL (1) | PL236550B1 (en) |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
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US20080213752A1 (en) * | 2004-11-01 | 2008-09-04 | Strategic Diagnostics Inc. | Bacteriophages as Selective Agents |
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2017
- 2017-12-11 PL PL423812A patent/PL236550B1/en unknown
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20080213752A1 (en) * | 2004-11-01 | 2008-09-04 | Strategic Diagnostics Inc. | Bacteriophages as Selective Agents |
Non-Patent Citations (3)
Title |
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CHEN J., BACTERIOPHAGE-BASED NANOPROBES FOR RAPID BACTERIA SEPARATION, 21 October 2015 (2015-10-21) * |
CHEN J., DETECTION OF ESCHERICHIA COLI IN DRINKING WATER USING T7 BACTERIOPHAGE-CONJUGATED MAGNETIC PROBE, 1 September 2015 (2015-09-01) * |
LIANA A.E., T4 BACTERIOPHAGE CONJUGATED MAGNETIC PARTICLES FOR E. COLI CAPTURING: INFLUENCE OF BACTERIOPHAGE LOADING, TEMPERATURE AND TRYPTONE, 1 March 2017 (2017-03-01) * |
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Publication number | Publication date |
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PL236550B1 (en) | 2021-01-25 |
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