# US 5,348,862 A · Method for the rapid determination of the microorganism content of liquids and fluid solutions, and apparatus for implementing this method

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 5,348,862 A
- Document ID: US5348862A
- Status: Granted
- Kind: A
- Application number: 07/947,477
- Filed: 1992-09-21
- Granted: 1994-09-20
- Earliest claimed: 1991-11-20
- Assignees: Biosensori S.p.A.
- Inventors: Edoardo Pasero; Carlo Rossetti; Franco Aiolfi
- Web page: https://patentgrep.com/patent/US5348862A

## CPC

- C12Q 1/04: Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor {(C12Q1/6897 takes precedence)}
- C12Q 1/06: Quantitative determination
- Y10S 435/817: Enzyme or microbe electrode
- Y10S 436/806: Electrical property or magnetic property
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

Method for amperiometrically measuring the bacterial content of liquids in a bioelectrochemical cell wherein the mediator is initially oxidized in the cell, then brought into contact with a capturing system containing the bacteria of the sample, resulting in a first response at the electrodes, and finally brought into contact again with the capturing system after the bacteria present have been killed, producing a second response due to the interfering substances, which is subtracted from the first response. An apparatus is also provided for implementing this method.

## Claims

### Claim 1

Method for measuring the bacterial content of liquids using amperometric means in a bioelectrochemical cell comprising initially oxidizing an electron transfer mediator in the cell, then bringing the mediator into contact with a filter containing the bacteria of a liquid sample, to produce a first response at the electrodes, and finally bringing the mediator into contact again with the filter after the bacteria present have been killed by chemical means to produce a second response due to interfering substances, which is subtracted from the first response to produce data representative of the bacterial content of the sample.

### Claim 2 (depends on 1)

Method according to claim 1, wherein the mediator is dissolved in a saline solution containing all the necessary ions for keeping the bacteria alive.

### Claim 3 (depends on 1)

Method according to claim 1 the reference electrode of the bioelectrochemical cell is protected with a membrane pervious to small ions, but impervious to mediators used for the amperometric detection of the metabolic activity of bacteria.

### Claim 4 (depends on 1)

Method according to claim 1 wherein the working electrode of the bioelectrochemical cell consists of sintered graphite or of any other porous current-conducting material.

### Claim 5 (depends on 1)

Method according to claim 1 in which the sample to be analysed is filtered using filter which is subsequently washed and inserted in the measuring circuit after the mediator has been oxidised.

### Claim 6 (depends on 1)

Method according to claim 1, wherein the bacteria retained on the filter are subjected to incubation before insertion of the filter in the measuring circuit.

### Claim 7 (depends on 1)

Method for measuring the bacterial content of liquids using amperometric means in a bioelectrochemical cell according to claim 1 comprising the introduction of a solution containing the mediator into the cell circuit; circulation of the solution until the mediator has been completely oxidised; filtration of the sample via the filter; washing of the filter; insertion of the filter into the cell circuit; detection of a first amperometric response; further washing of the filter with a biocide; insertion of the filter into the circuit again; detection of a second amperometric response which is subtracted from the first response; and conversion of the resultant datum into the measurement of the bacterial content.

### Claim 8 (depends on 1)

Device for implementing the method according to claim 1 comprising a bioelectrochemical cell; a first flow circuit which connects an outlet of the cell to an inlet of the cell; means for circulating a mediator liquid in this circuit; a second flow circuit in parallel with said first flow circuit which connects the outlet of the cell to the inlet of filter, and the outlet from this filter to the inlet of the cell; valve means for alternately bringing into circulation said first or said second flow circuit; a storage vessel for a mediator liquid designed to communicate with said first flow circuit so as to supply said first flow circuit with a quantity of mediator solution; a storage vessel for a biocidal liquid designed to communicate with said filter so as to cause a biocidal liquid to circulate therein; and a data processing circuit connected to the output of the electrodes of the bioelectrochemical cell for processing the amperometric data supplied by said electrodes.