CN1896269A - Identification and fast medicinal hypersensition determination for common clinical germ - Google Patents
Identification and fast medicinal hypersensition determination for common clinical germ Download PDFInfo
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- CN1896269A CN1896269A CN 200610045024 CN200610045024A CN1896269A CN 1896269 A CN1896269 A CN 1896269A CN 200610045024 CN200610045024 CN 200610045024 CN 200610045024 A CN200610045024 A CN 200610045024A CN 1896269 A CN1896269 A CN 1896269A
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Abstract
This invention discloses a process for simple identification and rapid susceptibility test of clinical common pathogenic bacteria. This test process includes the following steps: primary identification of gram-positive bacillus, gram-positive coccus, gram-negative bacilus and gram-negative diplococcus by direct observation of colonial morphology; identification of oxidative and fermentive bacteria by oxidase test based on colony and thallus morphology, and different biochemical identification and susceptibility test; or taking susceptibility test before identification and determining range of antibiotics which will provide the clinician with the drug susceptibility result before the identification of pathogen. In this process the result will be determined 1to 2 days earlier than traditional susceptibility test, which will save time, lighten the burden of sufferers and meet the needs of clinical treatment better.
Description
(1) technical field under
The present invention relates to a kind of simple Identification and susceptibility rapid assay methods of clinical common pathogenic bacteria, is culture of isolated, evaluation and drug sensitive detection are carried out in the hospital clinical check to the pathogenic bacteria of infectious diseases a kind of laboratory simple diagnostic method.
(2) background technology
China's hospital clinical microorganism modernization authenticate technology is subjected to the control of foreign country always, and semi-automatic microorganism identification equipment of import also will spend Renminbi hundreds of thousands of unit, and the reagent costliness, the unable use of middle and small hospital.Traditional authentication method, the tired rope of operation, poor accuracy.Many middle and small hospitals have stopped this inspection routine for this reason, the medicine waste that causes the doctor to write a prescription by rule of thumb to be brought, and the patient increases one's load and resistance.
(3) summary of the invention
The present invention provides a kind of evaluation and susceptibility rapid assay methods of clinical common pathogenic bacteria in order to remedy the deficiencies in the prior art, provides convenient diagnoses and treatment foundation by this simple and easy rapid identification method and drug sensitive detection result, for the clinician.
The present invention is achieved through the following technical solutions:
A kind of evaluation of clinical common pathogenic bacteria and susceptibility rapid assay methods, its special character is: comprise the steps:
(1) but by observing the colonial morphology preliminary evaluation be: Gram-positive bacillus, gram-positive cocci, gram negative bacillus, gram-negative diplococci;
(2) carry out oxidase test according to bacterium colony and thalli morphology and can be divided into oxidation bacterium and zymophyte, select different biochemical identification method program and drug sensitive test then;
(3) do drug sensitive test in the time of to above Bacteria Identification, at first dominant bacteria is carried out bacterium colony and observe, if the existence of single bacterium colony, desirable single bacterium colony smear staining microscopy are arranged, according to colonial morphology and thalli morphology, gram-positive cocci or gram negative bacillus are selected the microbiotic scope;
(4) get an identical 4-5 bacterium colony and do drug sensitive test;
(5) can provide the medication foundation to the clinician in advance, before strain identification, earlier drug sensitivity tests be informed the clinician, point out clinical gram-positive cocci or the gram negative bacillus of being separated into simultaneously.
The present invention is the classical summary of experience under the microbiology basic theories instructs.But by observing the colonial morphology preliminary evaluation be: Gram-positive bacillus, gram-positive cocci (staphylococcus, suis, faecalis), gram negative bacillus, gram-negative diplococci.According to bacterium colony and thalli morphology.Carry out oxidase test and can be divided into oxidation bacterium (oxidase positive) and zymophyte (oxidase negative), select different biochemical identification method program and drug sensitive test then.
(A) simple Identification method
1, enterobacteriaceae lactobacteriaceae and acinetobacter (oxidase negative)
The encountered pathogenic bacteria of enterobacteriaceae lactobacteriaceae and acinetobacter can be identified kind with 14 biochemical tests.
2, Rhodopseudomonas and other oxidase positive bacterium (oxidase positive)
The Bacteria Identification of Rhodopseudomonas can be arrived kind with 13 biochemical tests.
3, staphylococcus
Can determine clinical modal 8 kinds of staphylococcuses by 6 biochemical reactions.
4, streptococcus
(1) β Streptococcus hemolyticus: can identify by 4 biochemical reactions
(2) non-β Streptococcus hemolyticus: can identify by 5 biochemical reactions
5, faecalis
Can adopt 6 kinds of biochemical reaction pipes to be identified
6, dysentery bacterium in the stool sample and Corynebacterium diphtheriae
With 3 biochemical reaction items, identified with the factor serum aggegation
7, the evaluation of characteristic bacterium
The characteristic bacterium is meant the bacterium that can tentatively judge from bacterium colony and thalli morphology.Just can determine bacteria name by few distinctive biochemical identification test then.
(1) streptococcus pneumoniae: available 2 biochemical reactions are identified.
(2) streptococcus aureus: available 2 biochemical reactions are identified.
(3) Pseudomonas aeruginosa: available 2 biochemical reactions are identified.
(4) Shewanella putrefaciens: available 2 biochemical reactions are identified.
(5) faecalis: available 2 biochemical reactions are identified.
(6) micrococcus sp: available 2 biochemical reactions are identified.
(7) gemella haemolysans: available 2 biochemical reactions are identified.
(8) catarrh Blanc Chinese bacterium: available 2 biochemical reactions are identified.
(9) stenotrophomonas maltophilia: available 3 biochemical reactions are identified.
(10) Vibrio: available 3 biochemical reactions are identified.
(11) common distortion and Proteus mirabilis: available 3 biochemical reactions are identified.
(12) root Salmonella not: available 3 biochemical reactions are identified.
(13) listeria: available 2 biochemical reactions are identified.
(14) Aeromonas: available 3 biochemical reactions are identified.
(15) star nocardia: available 2 biochemical reactions are identified.
(16) non-tuberculous mycobacteria: bacterium colony and thalli morphology just can be identified.
(17) Hemolytic streptococcus (A, B group): available 2 biochemical reactions are identified.
(B) quick medicine-sensitive method:
To doing drug sensitive test before the above Bacteria Identification.At first dominant bacteria is carried out bacterium colony and observe, if the existence of single bacterium colony is arranged, desirable single bacterium colony smear staining microscopy, according to colonial morphology and thalli morphology, gram-positive cocci or gram negative bacillus are selected the microbiotic scope.Get identical bacterium colony and do drug sensitive test more than 4.It is dull and stereotyped to get the pure branch of bacterium colony simultaneously, uses in order to doing the bacterium biochemical reaction next day.Can provide to the clinician in advance and use the susceptibility foundation.Before strain identification, earlier drug sensitivity tests is informed the clinician, point out clinical gram-positive cocci or the gram negative bacillus of being separated into simultaneously.(this method must possess single bacterium colony more than 4 is arranged on the flat board).
1, gram-positive cocci can select for use following 14 kinds of different microbiotic to do drug sensitive test:
Penicillin, penbritin, cefoxitin, vancomycin, erythromycin, Cephazolin, cefuroxime, cefepime, Ofloxacine USP 23, levofloxacin, gentamicin, imipenum, trimethoprim-sulfamethoxazole, rifampin.
2, gram negative bacillus can select for use following 14 kinds of different microbiotic to do drug sensitive test:
Penbritin, oxygen piperazine Qin penicillin, peace Qu Nan, cefuroxime, cefotaxime, ceftazime, cefepime, cefoperazone/Sulbactam, imipenum, meropenem, amikacin, gentamicin, levofloxacin, trimethoprim-sulfamethoxazole.
3, faecalis can be selected following 7 kinds of microbiotic for use:
Penicillin, penbritin, gentamicin, vancomycin, Ciprofloxacin, levofloxacin, furadantin.
The present invention be the inventor in the working practice of decades, summed up simple Identification and the susceptibility rapid assay methods of this cover to pathogenic bacteria.This method does not need import equipment, is different from traditional authentication method yet, and sample is identified that accuracy rate can reach more than 90% equally, and easy and simple to handle, with low cost.Save a large amount of expenses not only for the middle and small hospital lab construction, and improved the quality of seeing a doctor of hospital, alleviated patient suffering and economical load.This solves masses' difficulty and high cost of getting medical for reducing resistance, and high cost of getting medical treatment problem all has crucial meaning.
To the evaluation of clinical common pathogenic bacteria, do not need specific installation, also be different from tradition coding authentication method, germ can be identified out with several distinctive biochemical reagents, the material-saving that saves time, quick and convenient, accurately stable; Special " drug sensitive test plate ", it is intuitively convenient to have than traditional method, fast advantage.Pathogenic bacterium do drug sensitive test earlier, and identify the back, has adapted to the needs of clinical diagnosis treatment more.This method shifts to an earlier date 1-2 days than traditional susceptibility test method(s) and goes out the result, saves time, and alleviates the every burden of patient, adapts to the needs of clinical treatment more.
(4) description of drawings
The invention will be further described below in conjunction with accompanying drawing.
Fig. 1 is common clinical pathogenic bacteria biochemical identification plate among the present invention;
Fig. 2 among the present invention to the drug sensitive detection plate of bacterium.
(5) embodiment
Embodiment 1: to the biochemical identification of pathogenic bacteria
This embodiment is to the biochemical identification program of pathogenic bacteria: (1) with ordinary method to the pathogenic bacteria sample cultivate, separation, purifying; (2) gramstaining, microscopy, observation form; (3) oxidase test is determined positive and negative; (4) biochemical identification is mixed with the bacteria suspension of 0.5-0.6 Maxwell unit's concentration with the pathogenic bacteria of separation and purification, and with the amount of pipettor by 150 μ l, the order by the biochemical cup of 1-14 item joins in each biochemical identification cup respectively, and identification plate is built; (5) identification plate is put into incubator, under 35 ℃ of conditions, cultivated 18-24 hour; (6) the biochemical identification plate that will cultivate is put into automatic identifying system and is identified, the title of pathogenic bacteria can be identified out less than half a minute.
Bacterium is differentiated example fast:
(1) streptococcus pneumoniae
Colonial morphology: recessed pleat (autolytic enzyme) in the middle of the α haemolysis grey small colonies, bacterium colony;
Thalli morphology: Gram-positive diplococcus, the two or catenation of many one-tenth;
Telling test: opeochin sensitivity, the bile bacteriolyze positive.
(2) catarrh Blanc Chinese bacterium
Colonial morphology: canescence or light yellow;
Thalli morphology: the Gram-positive diplococcus, be kidney shape, relatively other Neisserias are big for thalline, become double row more;
Telling test: DNA enzyme positive, the nitrate positive.
(3) Aeromonas
Colony characteristics: grow vigorous, bacterium colony is big and flat, β haemolysis is arranged (not haemolysis strain is also arranged);
Thalli morphology: gram negative bacillus, the hanging drop microscopy is active;
Telling test: oxidase positive, 6.0%Nacl does not grow, 0/129 resistance.
Fig. 1 is 14 biochemical identification plates of common clinical pathogenic bacteria among the present invention.Among the figure, the 1-14 on top is a sequence number (SN), and the circle of bottom is represented biochemical cup and the culture substrate of cultivating, the outer representative of circle biochemical culturing rack with cover.
Embodiment 2: to the determination of drug sensitivity method of pathogenic bacteria
This embodiment is to the determination of drug sensitivity program of pathogenic bacteria: (1) makes drug sensitive detection (needing 4-5 identical bacterium colony) when dividing pure pathogenic bacteria.(2) be mixed with the bacteria suspension of 0.5-0.6 Maxwell unit concentration, be coated on equably with cotton swab on the substratum of the M-H drug sensitive detection plate of having got ready; (3) after the compacting, build lid at the center of pasting each grid in " drug sensitive detection plate " respectively according to 14 kinds of antibiotic scraps of paper of gram-positive cocci (or gram negative bacillus) in the above quick medicine-sensitive method; (4) put into incubator, under 35 ℃ of conditions, cultivated 18-24 hour; (5) observations is analyzed the sensitivity of various antibiotic, determines the drug sensitive detection result.
Fig. 2 among the present invention to 14 kinds of drug sensitive detection plates of bacterium.Among the figure, the 1-14 on top is a sequence number (SN), and the circle of bottom is represented drug sensitive test paper, and circle is outer to be represented substratum and inoculate bacterium, and left side portion and sequence number (SN) 1 and sequence number (SN) 8 downsides are the susceptibility scale.
Claims (1)
1. the evaluation of a clinical common pathogenic bacteria and susceptibility rapid assay methods is characterized in that: comprise the steps:
(1) but by observing the colonial morphology preliminary evaluation be: Gram-positive bacillus, gram-positive cocci, gram negative bacillus, gram-negative diplococci;
(2) carry out oxidase test according to bacterium colony and thalli morphology and can be divided into oxidation bacterium and zymophyte, select different biochemical identification method program and drug sensitive test then;
(3) to doing drug sensitive test before the above Bacteria Identification, at first dominant bacteria is carried out bacterium colony and observe, if the existence of single bacterium colony, desirable single bacterium colony smear staining microscopy are arranged, according to colonial morphology and thalli morphology, gram-positive cocci or gram negative bacillus are selected the microbiotic scope;
(4) get 4 identical above bacterium colonies and do drug sensitive test;
(5) can provide the medication foundation to the clinician in advance, before strain identification, earlier drug sensitivity tests be informed the clinician, point out clinical gram-positive cocci or the gram negative bacillus of being separated into simultaneously.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101469340B (en) * | 2007-12-26 | 2012-05-02 | 上海复星佰珞生物技术有限公司 | Identification plate for gram positive aerobic bacteria and preparation thereof |
| CN106350571A (en) * | 2016-08-30 | 2017-01-25 | 北京市农林科学院 | Method for rapidly screening sensitive antibiotic for treating cow mastitis |
| CN108531542A (en) * | 2018-03-19 | 2018-09-14 | 中国水产科学研究院黄海水产研究所 | Automatic drug sensitivity test device for gram-negative bacteria and application method |
| CN110004205A (en) * | 2019-03-01 | 2019-07-12 | 昆明金域医学检验所有限公司 | The automatic audit method for early warning of microorganism medicine sensitive test, device and readable storage medium storing program for executing |
| CN114686556A (en) * | 2020-12-31 | 2022-07-01 | 石家庄市疾病预防控制中心（石家庄市卫生检测中心） | Method for detecting flora in stomach |
-
2006
- 2006-06-23 CN CN 200610045024 patent/CN1896269A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101469340B (en) * | 2007-12-26 | 2012-05-02 | 上海复星佰珞生物技术有限公司 | Identification plate for gram positive aerobic bacteria and preparation thereof |
| CN106350571A (en) * | 2016-08-30 | 2017-01-25 | 北京市农林科学院 | Method for rapidly screening sensitive antibiotic for treating cow mastitis |
| CN106350571B (en) * | 2016-08-30 | 2019-09-24 | 北京市农林科学院 | A kind of method of the sensitive antibiotics of quick screening treatment mastitis for milk cows |
| CN108531542A (en) * | 2018-03-19 | 2018-09-14 | 中国水产科学研究院黄海水产研究所 | Automatic drug sensitivity test device for gram-negative bacteria and application method |
| CN108531542B (en) * | 2018-03-19 | 2021-11-26 | 中国水产科学研究院黄海水产研究所 | Automatic drug sensitivity test device for gram-negative bacteria and application method |
| CN110004205A (en) * | 2019-03-01 | 2019-07-12 | 昆明金域医学检验所有限公司 | The automatic audit method for early warning of microorganism medicine sensitive test, device and readable storage medium storing program for executing |
| CN114686556A (en) * | 2020-12-31 | 2022-07-01 | 石家庄市疾病预防控制中心（石家庄市卫生检测中心） | Method for detecting flora in stomach |
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