CN105483207A - Rapid numerical identification method for enterobacteria of food-borne pathogenic bacteria - Google Patents
Rapid numerical identification method for enterobacteria of food-borne pathogenic bacteria Download PDFInfo
- Publication number
- CN105483207A CN105483207A CN201510866404.3A CN201510866404A CN105483207A CN 105483207 A CN105483207 A CN 105483207A CN 201510866404 A CN201510866404 A CN 201510866404A CN 105483207 A CN105483207 A CN 105483207A
- Authority
- CN
- China
- Prior art keywords
- biochemical
- qualification
- taxon
- bacterium
- test
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/10—Enterobacteria
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2304/00—Chemical means of detecting microorganisms
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a rapid numerical identification method for enterobacteria of food-borne pathogenic bacteria. The rapid numerical identification method includes confirming enterobacteria types and positive probabilities; selecting biochemical reaction required in a numerical method; establishing an enterobacteria database and determining a calculating method in the numerical method; detecting and identifying the bacteria to be detected. The rapid numerical identification method for the enterobacteria of the food-borne pathogenic bacteria has the advantages that pure cultures can be subjected to Gram staining, oxidase and OF experiment in advance, and then the rapid numerical identification method is adopted, so that a satisfactory identification result on the enterobacteria can be obtained; by the rapid numerical identification method, 139 bacteria of 31 categories and 8 unclassified floras, namely 159 classified units in total of enterobacteriaceae can be identified, and Yersinia enterocolitica can be identified to 6 kinds of biochemical types; the rapid numerical identification method is much greater in identified bacteria quantity and much higher in identification accuracy than two existing internationally-recognized similar identification methods on the market.
Description
Technical field:
The invention belongs to biochemical identification technical field, be specifically related to the numerical value rapid identification method of a kind of food-borne pathogens Chang Gan section bacterial classification.
Background technology:
Food safety is the significant problem being related to human health and national economy, and microbial safety is the most important thing of food safety.Owing to containing abundant nutritive substance in food, after microbial contamination food, Fast-propagation quantity is that geometry level increases, thus causes serious food-safety problem.According to statistics, there is food poisoning 6685 in 2012 in China, wherein microbes food poisoning Poisoning Number is maximum, accounts for 56.1% of sum; 1988-2008, American Electronic food borne outbreak reporting system (eFORS) reports 13405 food origin diseases altogether and breaks out event, and wherein 3613 food borne outbreak events have bacterium to cause, and account for that known cause of disease breaks out event 45%.Enterobacteriaceae lactobacteriaceae distribution is wide, and existing sorting technique is divided into 42 genus and 8 non-classified groups, comprises Salmonellas, Shigellae and Enteropathogenic Escherichia coli, Yersinia enterocolitica etc. and breaks out relevant important pathogenic bacteria to food origin disease.Therefore be a main task of inspection and quarantine Microbiological Lab to the screening and identification of Chang Gan section bacterium.The research-and-development activity of this research team long campaigns food microorganisms security fields, have developed serial Fast Detection Technique, preservation of bacteria strain 20000 strain altogether at present in food-borne pathogens microorganism resource, wherein Chang Gan section bacterium is more than 10000 strains, for the research of all kinds of Bacteria Detection technology provides resource base.
Bacteria screening is biochemical identification method with the most widely used general most basic method of qualification.The qualification of bacterium numerical value is in conjunction with the biochemical identification method that digital appraisal principle develops on the basis of biochemical identification method, this method has the advantage such as directly perceived, reliable of conventional biochemical identification method, combine Digital identification principle simultaneously, make qualification result have more science and repeatability.
Chang Gan section bacterium is that Gram-negative is shaft-like, the bacterium of fermented type, Zhousheng and oxidase negative.It all has special importance on medical science, agronomy and genetics research.Therefore, people pay attention to greatly.Although Chang Gan section bacterium is comparatively clear and definite with the boundary line of contiguous Cordycepps, do not have boundary line clearly between belonging to, therefore, they are not easily differentiated each other.The biochemical identification of current Chang Gan section bacterium mainly contains traditional substratum back-and-forth method, micro biochemical pipe identification method and numerical value biochemical identification method.First two method wastes time and energy, numerical value identification method is quick with it, identification rate is high, be convenient to realize the advantages such as automatization, commercialization, stdn, now become the predominant methods of food sanitation and the field Bacteria Identification such as clinical, made it have been widely recognized at world wide and develop fast.
Current commercial Chang Gan section numerical value identification systems have the API20E of the French Mei Liai company being described as " global gold standard ", first identification systems be developed, without the need to instrumentation, stdn manual method completes the biochemical identification of bacterium, for low cost substitutes the best product of traditional-handwork bacterial biochemical assay.API20E entero-bacte indentifying substance bar identified gram negative bacillus in 24 hours, cover clinical common bacterium, comprise 70 Ge Chang bar section's bacterial classifications and 32 other Intestinal floras, such as oxidase positive zymophyte-3 Aeromonass, 6 Vibrios, 3 Pasteurellas, non-zymocyte-1 acinetobacter, 4 Rhodopseudomonass etc.And Chang Gan section mycorhiza is according to 2 times more than of the nearly above-mentioned numerical value of uncle's Jie Shi handbook.Therefore the taxon quantity that contains of API20E is relatively less.The MID-GNA of the comparatively popular Microgen company of numerical value identification systems Ze Shi Britain accepted of another money, or/and MID-GNB, MID-GNA can be used alone, can identify 41 Ge Chang bar section's bacterial classifications and 3 non-zymocyte acinetobacters; MID-GNA and MID-GNB with the use of 134 Ge Chang bar section bacterium and 41 other Gram-negative bacterias can be identified, such as non-zymocyte-1 acinetobacter, 7 Rhodopseudomonass, oxidase positive zymophyte-11 Vibrios, 4 Aeromonass, 2 Pasteurellas etc.Although the taxon quantity that the product of Microgen company is contained is many compared with API20E, say with regard to accuracy, user is more prone to API20E.The database of biochemical identification system is the heart of whole system, basic data and positive probability misapplication, and qualification result error rate can be made to raise.We find, in database there is certain difference in Citrobacteramalonaticus (without malonate citric acid bacillus) API and MID system on urine enzyme, the positive probability of occurrence that API urinates enzyme is 1%, MID is then 85%, the reason occurring this situation mainly the positive probability that occurs of biochemical test and the method for inspection closely related, as urea, hydrogen sulfide etc., the frequency that different methods occurs is different, therefore causes qualification result inconsistent.
Summary of the invention:
The object of the invention is the shortcoming existed for numerical value authentication method in prior art, provide a kind of more comprehensively, the numerical value rapid identification method of food-borne pathogens Chang Gan section bacterial classification more accurately.
The numerical value rapid identification method of food-borne pathogens Chang Gan section of the present invention bacterial classification, is characterized in that, comprise the following steps:
The probability that a, the kind determining Chang Gan section bacterium and at least 24 biochemical tests appearance thereof are positive, 24 described biochemical tests are: phenylalanine deamination, wood sugar, raffinose, indoles, sucrose, ornithine decarboxylase, lactose, ONPG, urine enzyme, Citrate trianion, maltose, lysine decarboxylase, malonate, sorbyl alcohol, galactitol, pectinose, melibiose, rhamnosyl, Vitamin C2, MR, H
2s, trehalose, cellobiose, oxydase; The positive probability of these biochemical tests can according to authoritative data such as uncle Jie Shi handbooks, and obtaining each biochemical test occurs positive probability;
24 biochemical tests in b, selection step a are as biochemical reaction required in numerical method:
C, set up the database of Chang Gan section bacterium, determine numerical method Computational Methods: according to the positive probability of the taxon selected and biochemical test, form a probability matrix;
Determine the method for calculation of total frequency of occurrences of each taxon, multinomial total occurrence frequency, qualification percentage ratio according to bacterium probability appraisal theory, specific formula for calculation is as follows:
Total frequency of occurrences of each taxon equals the product of the positive probability of each biochemical test corresponding to this taxon;
Multinomial total occurrence frequency equals the summation of total frequency of occurrences of each taxon;
Qualification percentage ratio equals the total frequency of occurrences of each taxon and is multiplied by 100 again divided by multinomial total occurrence frequency, and all qualification percentage ratio sums should equal 100;
After d, test strains pure culture, carry out gramstaining, oxydase, OF tests, gramstaining is selected to be negative, oxidase negative, OF is 24 kinds of biochemical tests that the bacterial strain of fermented type carries out step a, can the qualification bar of commodity in use, also can buy biochemical tube or prepare substratum voluntarily and carry out biochemical test;
After 24 kinds of biochemical test results of e, acquisition test strains, according to yin, yang, the unknown-value of biochemical reaction, according to method of calculation, taxons all in database are calculated, first biochemical test value is converted to probable value, i.e. the positive probability value of experimental result to be the positive be exactly then corresponding biochemical test; If feminine gender is the 100 positive probability values deducting biochemical test then, if the above results 0 or 100, then replace by approximation 1 or 99, again according to the method for calculation of step c calculate each taxon total frequency of occurrences, multinomial total occurrence frequency, identify that if percentage ratio a certain biochemical test result is for unknown or smudgy, then this biochemical test item does not participate in calculating, thus does not affect final qualification result;
F, according to calculation result, to judge identification of strains result to be measured: the size according to qualification percentage ratio sorts, the highest corresponding bacterial strain of qualification percentage ratio is the taxon belonging to test strains.The formulation of judgement criteria is on the basis of existing commercial two kinds of identification systems of generally acknowledging in the world, carry out in conjunction with Chinese Shi Yuanxingchang bar section bacterial classification main biochemical type, serotype, genotype aspect, being with Chinese characteristics property, is more suitable for the qualification result evaluation of domestic enterobacteriaceae bacterium than two kinds of identification systems of generally acknowledging.
Preferably, inconsistent biochemical test is all provided to the qualification of test strains and out single-row.
Preferably, in result display, provide Chinese to taxon, Latin contrasts, and meets Chinese writing style.
Of the present invention determine enterobacteriaceae 31 belong to 139 kinds, amount to the positive probability of 159 taxons and 47 kinds of (comprising above-mentioned 24 kinds of biochemical tests) biochemical tests thereof, these 159 taxons are as follows respectively: 1-Escherichiaspp (Escherichia), 2-Escherichiacoli (colon bacillus), 3-Escherichiacol, inactive (colon bacillus, torpescence), 4-Escherichiablattae (cockroach Escherichia), 5-Escherichiafergusanii (Cathy Ferguson Escherichia), 6-Escherichiahermannii (He Shi Escherichia), 7-Escherichiavulneris (wound Escherichia), 8-Budviciaaquatia (aquatic Boot ties up western bacterium), 9-Buttiauxellaspp (cloth mound Bacillaceae), 10-Buttiauxellaagrestis (country's cloth mound bacillus), 11-Buttiauxellabrennerae, 12-Buttiauxellaferragutiae, 13-Buttiauxellagaviniae, 14-Buttiauxellaizardii, 15-Buttiauxellanoackiae (Nuo Shi cloth mound bacillus), 16-Buttiauxellawarmboldiae, 17-cedeceaspp (Cedecea), 18-cedeceadavisae (Cedecea davisae), 19-cedecealapagei (Cedecea lapagei), 20-cedeceaneteri (Cedecea neteri), 21-cedeceaspecies3 (westernly western bacterium No. 3), 22-cedeceaspecies5 (westernly western bacterium No. 5), 23-Citrobacterspp (Citrobacter), 24-Citrobacteramalonaticus (without malonate citric acid bacillus), 25-Citrobacterfreundii (citrobacter freundii), 26-Citrobacterbraakii (Bu Shi citric acid bacillus), 27-Citrobacterfarmeri (Fa Shi citric acid bacillus), 28-Citrobactergillenii, 29-Citrobacterkoseri (C.diversus) (difference citric acid bacillus), 30-Citrobactermurliniae, 31-Citrobacterrodentium (corrosion citric acid bacillus), 32-Citrobactersedlakii (Sai Shi citric acid bacillus), 33-Citrobacterwerkmanii (Wei Shi citric acid bacillus), 34-Citrobacteryoungae (Young citric acid bacillus)), 35-Edwardsiellaspp (Edwardsiella), 36-Edwardsiellatarda (Edwardsiella tarda), 37-Edwardsiellatardabiogroup1 (edwardsiella tarda biotype 1), 38-Edwardsiellahoshinae (guarantor section tarda), 39-Edwardsiellaictaluri (Edwardsiella ictaluri), 40-Enterobacterspp (enterobacter), 41-Enterobactercloacae (enterobacter cloacae), 42-Enterobacteraerogenes (enteroaerogen), 43-Enterobacteramnigenusbiogroup1 (Enterobacter amnigenus biotype 1), 44-Enterobacteramnigenusbiogroup2 (Enterobacter amnigenus biotype 2), 45-Enterobacterasburiae (A Sibu enterobacteria), 46-Enterobactercancerogenus (E.taylorae) (raw cancer enterobacteria), 47-Enterobacterdissolvens (enterobacter dissolvens), 48-Enterobactergergoviae (Ge Gaofei enterobacteria), 49-Enterobacterhormaechei (Huo meter Qi enterobacteria), 50-Enterobacterintermedius (enterobacter intermedium), 51-Enterobacternimipressuralis (Enterobacter nimipressuralis), 52-Enterobacterpyrinus (pyriform enterobacteria), 53-Enterobactersakazakii (E.sakazakii), 54-Enterobacteragglomeransgroup enterobacter agglomerans, the raw cancer enterobacteria of 55-Enterobactercancerogenus, 56-Ewingellaamericana (Wen bacterium is liked in America), 57-Hafniaspp. (Hafnia), 58-Hafniaalvei (hafnia alvei), 59-Hafniaalveibiogroup1 (biological 1 group of hafnia alvei), 60-Klebsiellaspp (klebsiella spp), 61-Klebsiellapneumoniaesubsp.Ozaenae (Klebsiella ozaenae), 62-Klebsiellapneumoniaesubsp.Pneumoniae (Klebsiella Pneumoniae), 63-Klebsiellapneumoniaesubsp.Rhinoscleromatis (Klebsiella rhinoscleromatis), 64-Klebsiellaoxytoca (Klebsiella oxytoca), 65-Klebsiellaoxytocaorithinepositive (Klebsiella ornithinolytica), 66-Klebsiellaplanticola (planting raw klebsiella), 67-Klebsiellaterrigena (kluyvera terrigena), 68-Kluyveraspp (Kluyvera), 69-Kluyveraascorbata (Kluyvera ascorbata), 70-Kluyveracryocrescens (Kluyvera cryocrescens), 71-Kluyveragenrgiana (Ke Lv Wal, Georgia bacterium), 72-Leclerciaadecarboxylata (Leclercia adecarboxylata), 73-Leminorellaspp (Leminorella), 74-Leminorellagrimontii (Green covers and strangles minot bacterium), 75-Leminorellarichardii (Richard Le meter Nuo bacterium), 76-Moellerellawisconsensis (Wisconsin Moeller bacterium), 77-Morganellaspp (Morganella), 78-Morganellamorganiisubsp.Morganii (Mo Genmo root fungus rub root subspecies), 79-Morganellamorganiisubsp.Sibonii (Mo Genmo root fungus Xi Baini subspecies), 80-Morganellamorganiibiogroup1 (biological 1 group of Mo Genmo root fungus), 81-Obesumbacteriumproteusbiogroup2 (obesumbacterium proteus, biological 2 groups), 82-Pantoeaspp (general Pseudomonas), 83-Pantoeaagglomerans, 84-Pantoeadispersa (disperseing general bacterium), 85-Photorhabdus (Photorhabdus), 86-Photorhabdusluminescens (luminous polished rod bacterium), 87-PhotorhabdusDNAhybridizationgroup5 (Photorhabdusasymbiotia) (non-co-third contact of a total solar or lunar eclipse bacillus), 88-Pragiafontium (Quan Ju Prague bacterium), 89-Proteusspp (proteus), 90-Proteusvulgaris (proteus vulgaris), 91-Proteusmirabilis (Proteus mirabilis), 92-Proteusmyxofaciens (producing mucus Bacillus proteus), 93-Proteuspenneri (Buddhist nun Pan Bacillus proteus), 94-Providenciaspp (Providencia), 95-Providenciaalcalifaciens (producing alkali Providence), 96-Providenciaheimbachae (Providencia heimbachae), 97-Providenciarettgeri (providencia rettgeri), 98-Providenciarustigianii (Providencia rustigianii), 99-Providenciastuarlii (providencia stuartii), 100-Rahnellaaquatilis (aquatic draw grace bacterium), 101-Salmonellaspp (salmonella), 102-Salmonellabongori (Bang Geer Salmonellas), 103-Salmonellaentericasubsp.Enterica (Salmonella enteritidis I), 104-Salmonellaentericasubsp.arizonae (Arizona Salmonellas III a), 105-Salmonellaentericasubsp.diarizonae (two-phase Arizona Salmonellas III b), 106-Salmonellaentericasubsp.houtenae (a person of outstanding talent Salmonellas IV), 107-Salmonellaentericasubsp.indica (India Salmonellas V), 108-Salmonellaentericasubsp.salamae (salam Salmonellas II), 109-Salmonellaserovarcholeraesuis (Salmonella choleraesuls I), 110-SalmonellaserovarGallinarum (avian infectious bronchitis nephritis virus I), 111-SalmonellaserovarparatyphiA (Salmonella paratyphi A I), 112-Salmonellaserovarpullorum (S. pullonum I), 113-Salmonellaserovartyphi (salmonella typhi I), 114-Serratiaspp (serratia), 115-Serratiamarcescens (serratia marcescens), 116-Serratiaentomophila (Serratia entomophil), 117-Serratiaficaria (serratia ficaria), 119-Serratialiquefaciens (liquefied Serratia), 120-Serratiamarcescensbiogroup1 (biological 1 group of serratia marcescens), 121-Serratiaodoriferabiogroup1 (biological 1 group of smell Serratia), 122-Serratiaodoriferabiogroup2 (biological 2 groups of smell Serratia), 123-Serratiaplymuthica (Plymouth Serratia), 124-Serratiarubidaea (serratia rubida), 125-Shigellaspp (Shigella), 126-Shigelladysenteriae (shigella dysenteriae, GroupA), 127-Shigellaboydii (Shigella bogdii, GroupC), 128-Shigellaflexneri (shigella flexneri, GroupB), 129-Shigellasonnei (Shigella sonnei, GroupD), 130-Tatumellaptyseos (phlegm Tatumella), 131-Trabulsiellaguamensis (Guam Te Labusi bacterium), 132-Xenorhabdusnematophilus (Xenorhabdus nematophilus), 133-Yersiniaspp (yersinia's genus), 134-Yersiniapestis (yersinia pestis), 135-Yersiniaaldovae (Ao Erdewa Yersinia), 136-Yersiniabercovieri (ripple is expected Yersinia), 137-Yersiniaenterocolitica (Yersinia enterocolitica), 138-Yersiniaenterocolitica1A (the biological 1A type of Yersinia enterocolitica), 139-Yersiniaenterocolitica1B (the biological 1B type of Yersinia enterocolitica), 140-Yersiniaenterocolitica2 (biological 2 types of Yersinia enterocolitica), 141-Yersiniaenterocolitica3 (biological 3 types of Yersinia enterocolitica), 142-Yersiniaenterocolitica4 (biological 4 types of Yersinia enterocolitica), 143-Yersiniaenterocolitica5 (biological 5 types of Yersinia enterocolitica), 144-Yersiniafrederiksenii (Yersinia fredericksenii), 145-Yersiniaintermedia (Yersinia intermedia), 146-Yersiniakristensenii (Yersinia kristensenii), 147-Yersiniamollaretii (Mo Lalai Yersinia), 148-Yersiniapseudotubercttlosis (yersinia pseudotuberculosis), 149-Yersiniarohdei (Luo Di Yersinia), 150-Yersiniaruckeri (Yersinia ruckeri), 151-Yokenellaregensburgei (Koserellatrabulsii) (Lei Jisibai receive York bacterium), 152-EntericGroup58, 153-EntericGroup59, 154-EntericGroup60, 155-EntericGroup63, 156-EntericGroup64, 157-EntericGroup68, 158-EntericGroup69, 159-EntericGroup137.
The advantage that present method has compared with additive method is:
1, the division bacteria in Chang Gan section bacterium is up-to-date, and bacterium adopts current up-to-date internal authority classification, and qualification result lists file names with Chinese and Latin title, meets Chinese custom.Current Chang Gan section bacterium is all listed by present method, false retrieval and undetected possibility greatly reduce, common enterobacteriaceae 31 can be identified and belong to 139 kinds and 8 non-classified groups, totally 159 taxons, and the Chang Gan section bacterium that internationally recognized standard identification systems API identifies its 1/2, MID Chang Gan section bacterium of identifying not enough is 126 kinds and 8 non-classified groups.
2, native system database is so that the basis of the authoritative data such as uncle's Jie Shi classification manual of latest edition and uncle Jie Shi identification handbook to build, and compared with API20E and MIDGNA+GNB, add the math block of the math block of each Pseudomonas, Yersinia enterocolitica kind 6 kinds of biotypes.
3, native system concludes out by bacteria bio response characteristic on the basis of uncle Jie Shi handbook, biochemical screening is reasonable, 24 biochemical reactions being different from API20E, MIDGNA+GNB filtered out, qualification result evaluation accurately and reliably, can by some dientification of bacteria that can not distinguish very well in above-mentioned two kinds of identification systems out:
(1) the qualification accuracy of the present invention to yersinia's genus is higher than API, MID, and Yersinia enterocolitica can be identified biotype, and API, MID then can only identify the level of Yersinia enterocolitica kind;
(2) API20E needs on the basis adding this biochemical test of yellow pigment, identify that percentage ratio just can reach more than 90% (after 21 biochemical tests in API20E system to the qualification of Enterobacter sakazakii, qualification percentage ratio is more than 50%), and native system directly just can obtain the result of more than 90% by 24 biochemical tests.
(3) the present invention identifies bar, the molecular biology PCR method of specific gene, 16SrDNA sequencing to Chang Gan section standard bacteria and actual sample through strict statistical analysis repeatedly and practice demonstration by API20E and GNA+GNB, optimizes the degree of confidence with confirmation present method.
Embodiment
Below by specific embodiment, the present invention is further detailed explanation, but embodiments of the present invention are not limited thereto.
The numerical value rapid identification method of food-borne pathogens Chang Gan section of the present invention bacterial classification
(1) kind and the positive probability thereof of Chang Gan section bacterium is determined
The present embodiment determination enterobacteriaceae 31 belongs to 139 kinds, amounts to the positive probability of 159 taxons and 47 kinds of (comprising above-mentioned 24 kinds of biochemical tests) biochemical tests thereof.These 159 taxons are as follows respectively: 1-Escherichiaspp (Escherichia), 2-Escherichiacoli (colon bacillus), 3-Escherichiacol, inactive (colon bacillus, torpescence), 4-Escherichiablattae (cockroach Escherichia), 5-Escherichiafergusanii (Cathy Ferguson Escherichia), 6-Escherichiahermannii (He Shi Escherichia), 7-Escherichiavulneris (wound Escherichia), 8-Budviciaaquatia (aquatic Boot ties up western bacterium), 9-Buttiauxellaspp (cloth mound Bacillaceae), 10-Buttiauxellaagrestis (country's cloth mound bacillus), 11-Buttiauxellabrennerae, 12-Buttiauxellaferragutiae, 13-Buttiauxellagaviniae, 14-Buttiauxellaizardii, 15-Buttiauxellanoackiae (Nuo Shi cloth mound bacillus), 16-Buttiauxellawarmboldiae, 17-cedeceaspp (Cedecea), 18-cedeceadavisae (Cedecea davisae), 19-cedecealapagei (Cedecea lapagei), 20-cedeceaneteri (Cedecea neteri), 21-cedeceaspecies3 (westernly western bacterium No. 3), 22-cedeceaspecies5 (westernly western bacterium No. 5), 23-Citrobacterspp (Citrobacter), 24-Citrobacteramalonaticus (without malonate citric acid bacillus), 25-Citrobacterfreundii (citrobacter freundii), 26-Citrobacterbraakii (Bu Shi citric acid bacillus), 27-Citrobacterfarmeri (Fa Shi citric acid bacillus), 28-Citrobactergillenii, 29-Citrobacterkoseri (C.diversus) (difference citric acid bacillus), 30-Citrobactermurliniae, 31-Citrobacterrodentium (corrosion citric acid bacillus), 32-Citrobactersedlakii (Sai Shi citric acid bacillus), 33-Citrobacterwerkmanii (Wei Shi citric acid bacillus), 34-Citrobacteryoungae (Young citric acid bacillus)), 35-Edwardsiellaspp (Edwardsiella), 36-Edwardsiellatarda (Edwardsiella tarda), 37-Edwardsiellatardabiogroup1 (edwardsiella tarda biotype 1), 38-Edwardsiellahoshinae (guarantor section tarda), 39-Edwardsiellaictaluri (Edwardsiella ictaluri), 40-Enterobacterspp (enterobacter), 41-Enterobactercloacae (enterobacter cloacae), 42-Enterobacteraerogenes (enteroaerogen), 43-Enterobacteramnigenusbiogroup1 (Enterobacter amnigenus biotype 1), 44-Enterobacteramnigenusbiogroup2 (Enterobacter amnigenus biotype 2), 45-Enterobacterasburiae (A Sibu enterobacteria), 46-Enterobactercancerogenus (E.taylorae) (raw cancer enterobacteria), 47-Enterobacterdissolvens (enterobacter dissolvens), 48-Enterobactergergoviae (Ge Gaofei enterobacteria), 49-Enterobacterhormaechei (Huo meter Qi enterobacteria), 50-Enterobacterintermedius (enterobacter intermedium), 51-Enterobacternimipressuralis (Enterobacter nimipressuralis), 52-Enterobacterpyrinus (pyriform enterobacteria), 53-Enterobactersakazakii (E.sakazakii), 54-Enterobacteragglomeransgroup enterobacter agglomerans, the raw cancer enterobacteria of 55-Enterobactercancerogenus, 56-Ewingellaamericana (Wen bacterium is liked in America), 57-Hafniaspp. (Hafnia), 58-Hafniaalvei (hafnia alvei), 59-Hafniaalveibiogroup1 (biological 1 group of hafnia alvei), 60-Klebsiellaspp (klebsiella spp), 61-Klebsiellapneumoniaesubsp.Ozaenae (Klebsiella ozaenae), 62-Klebsiellapneumoniaesubsp.Pneumoniae (Klebsiella Pneumoniae), 63-Klebsiellapneumoniaesubsp.Rhinoscleromatis (Klebsiella rhinoscleromatis), 64-Klebsiellaoxytoca (Klebsiella oxytoca), 65-Klebsiellaoxytocaorithinepositive (Klebsiella ornithinolytica), 66-Klebsiellaplanticola (planting raw klebsiella), 67-Klebsiellaterrigena (kluyvera terrigena), 68-Kluyveraspp (Kluyvera), 69-Kluyveraascorbata (Kluyvera ascorbata), 70-Kluyveracryocrescens (Kluyvera cryocrescens), 71-Kluyveragenrgiana (Ke Lv Wal, Georgia bacterium), 72-Leclerciaadecarboxylata (Leclercia adecarboxylata), 73-Leminorellaspp (Leminorella), 74-Leminorellagrimontii (Green covers and strangles minot bacterium), 75-Leminorellarichardii (Richard Le meter Nuo bacterium), 76-Moellerellawisconsensis (Wisconsin Moeller bacterium), 77-Morganellaspp (Morganella), 78-Morganellamorganiisubsp.Morganii (Mo Genmo root fungus rub root subspecies), 79-Morganellamorganiisubsp.Sibonii (Mo Genmo root fungus Xi Baini subspecies), 80-Morganellamorganiibiogroup1 (biological 1 group of Mo Genmo root fungus), 81-Obesumbacteriumproteusbiogroup2 (obesumbacterium proteus, biological 2 groups), 82-Pantoeaspp (general Pseudomonas), 83-Pantoeaagglomerans, 84-Pantoeadispersa (disperseing general bacterium), 85-Photorhabdus (Photorhabdus), 86-Photorhabdusluminescens (luminous polished rod bacterium), 87-PhotorhabdusDNAhybridizationgroup5 (Photorhabdusasymbiotia) (non-co-third contact of a total solar or lunar eclipse bacillus), 88-Pragiafontium (Quan Ju Prague bacterium), 89-Proteusspp (proteus), 90-Proteusvulgaris (proteus vulgaris), 91-Proteusmirabilis (Proteus mirabilis), 92-Proteusmyxofaciens (producing mucus Bacillus proteus), 93-Proteuspenneri (Buddhist nun Pan Bacillus proteus), 94-Providenciaspp (Providencia), 95-Providenciaalcalifaciens (producing alkali Providence), 96-Providenciaheimbachae (Providencia heimbachae), 97-Providenciarettgeri (providencia rettgeri), 98-Providenciarustigianii (Providencia rustigianii), 99-Providenciastuarlii (providencia stuartii), 100-Rahnellaaquatilis (aquatic draw grace bacterium), 101-Salmonellaspp (salmonella), 102-Salmonellabongori (Bang Geer Salmonellas), 103-Salmonellaentericasubsp.Enterica (Salmonella enteritidis I), 104-Salmonellaentericasubsp.arizonae (Arizona Salmonellas III a), 105-Salmonellaentericasubsp.diarizonae (two-phase Arizona Salmonellas III b), 106-Salmonellaentericasubsp.houtenae (a person of outstanding talent Salmonellas IV), 107-Salmonellaentericasubsp.indica (India Salmonellas V), 108-Salmonellaentericasubsp.salamae (salam Salmonellas II), 109-Salmonellaserovarcholeraesuis (Salmonella choleraesuls I), 110-SalmonellaserovarGallinarum (avian infectious bronchitis nephritis virus I), 111-SalmonellaserovarparatyphiA (Salmonella paratyphi A I), 112-Salmonellaserovarpullorum (S. pullonum I), 113-Salmonellaserovartyphi (salmonella typhi I), 114-Serratiaspp (serratia), 115-Serratiamarcescens (serratia marcescens), 116-Serratiaentomophila (Serratia entomophil), 117-Serratiaficaria (serratia ficaria), 119-Serratialiquefaciens (liquefied Serratia), 120-Serratiamarcescensbiogroup1 (biological 1 group of serratia marcescens), 121-Serratiaodoriferabiogroup1 (biological 1 group of smell Serratia), 122-Serratiaodoriferabiogroup2 (biological 2 groups of smell Serratia), 123-Serratiaplymuthica (Plymouth Serratia), 124-Serratiarubidaea (serratia rubida), 125-Shigellaspp (Shigella), 126-Shigelladysenteriae (shigella dysenteriae, GroupA), 127-Shigellaboydii (Shigella bogdii, GroupC), 128-Shigellaflexneri (shigella flexneri, GroupB), 129-Shigellasonnei (Shigella sonnei, GroupD), 130-Tatumellaptyseos (phlegm Tatumella), 131-Trabulsiellaguamensis (Guam Te Labusi bacterium), 132-Xenorhabdusnematophilus (Xenorhabdus nematophilus), 133-Yersiniaspp (yersinia's genus), 134-Yersiniapestis (yersinia pestis), 135-Yersiniaaldovae (Ao Erdewa Yersinia), 136-Yersiniabercovieri (ripple is expected Yersinia), 137-Yersiniaenterocolitica (Yersinia enterocolitica), 138-Yersiniaenterocolitica1A (the biological 1A type of Yersinia enterocolitica), 139-Yersiniaenterocolitica1B (the biological 1B type of Yersinia enterocolitica), 140-Yersiniaenterocolitica2 (biological 2 types of Yersinia enterocolitica), 141-Yersiniaenterocolitica3 (biological 3 types of Yersinia enterocolitica), 142-Yersiniaenterocolitica4 (biological 4 types of Yersinia enterocolitica), 143-Yersiniaenterocolitica5 (biological 5 types of Yersinia enterocolitica), 144-Yersiniafrederiksenii (Yersinia fredericksenii), 145-Yersiniaintermedia (Yersinia intermedia), 146-Yersiniakristensenii (Yersinia kristensenii), 147-Yersiniamollaretii (Mo Lalai Yersinia), 148-Yersiniapseudotubercttlosis (yersinia pseudotuberculosis), 149-Yersiniarohdei (Luo Di Yersinia), 150-Yersiniaruckeri (Yersinia ruckeri), 151-Yokenellaregensburgei (Koserellatrabulsii) (Lei Jisibai receive York bacterium), 152-EntericGroup58, 153-EntericGroup59, 154-EntericGroup60, 155-EntericGroup63, 156-EntericGroup64, 157-EntericGroup68, 158-EntericGroup69, 159-EntericGroup137.
(2) biochemical test required in numerical method is selected.According to the method setting up branching diagram, and the principle of maximum taxons can be distinguished with minimum biochemical test, select 24 biochemical tests.These 24 biochemical tests are as follows respectively: phenylalanine deamination, wood sugar, raffinose, indoles, sucrose, ornithine decarboxylase, lactose, ONPG, urine enzyme, Citrate trianion, maltose, lysine decarboxylase, malonate, sorbyl alcohol, galactitol, pectinose, melibiose, rhamnosyl, Vitamin C2, MR, H
2s, trehalose, cellobiose, oxydase.
(3) set up the database of Chang Gan section bacterium, determine numerical method Computational Methods.According to the taxon of selection and the positive probability of biochemical test, form a probability matrix.
The method of calculation of total frequency of occurrences of each taxon, multinomial total occurrence frequency, qualification percentage ratio are determined according to bacterium probability appraisal theory.
This calculation formula is as follows:
Total frequency of occurrences of each taxon equals the product of the positive probability of each biochemical test corresponding to this taxon;
Multinomial total occurrence frequency equals the summation of total frequency of occurrences of each taxon.
Qualification percentage ratio equals the total frequency of occurrences of each taxon and is multiplied by 100 again divided by multinomial total occurrence frequency, and all qualification percentage ratio sums should equal 100.
(4) after test strains pure culture, carry out gramstaining, oxydase, OF tests, select gramstaining to be negative, oxidase negative, OF is that the bacterial strain of fermented type carries out above-mentioned 24 kinds of biochemical tests (phenylalanine deamination, wood sugar, raffinose, indoles, sucrose, ornithine decarboxylase, lactose, ONPG, urine enzyme, Citrate trianion, maltose, lysine decarboxylase, malonate, sorbyl alcohol, galactitol, pectinose, melibiose, rhamnosyl, Vitamin C2, MR, H
2s, trehalose, cellobiose, oxydase).Can the qualification bar of commodity in use, also can buy biochemical tube or prepare substratum voluntarily and carry out biochemical test.
(5), after obtaining 24 kinds of biochemical test results, according to yin, yang, the unknown-value of biochemical reaction, according to method of calculation, taxons all in database are calculated.First biochemical values is converted to probable value, i.e. the positive probability value of experimental result to be the positive be exactly then corresponding biochemical test; If feminine gender is the 100 positive probability values deducting biochemical test then.If the above results 0 or 100, then replace by approximation 1 or 99.Total frequency of occurrences of each taxon, multinomial total occurrence frequency, qualification percentage ratio is calculated again according to the method for calculation of step (3), if a certain biochemical test result is unknown or smudgy, then this biochemical test item does not participate in calculating, thus does not affect final qualification result;
(6) according to calculation result, identification of strains result to be measured is judged.Size according to qualification percentage ratio sorts, and the highest corresponding bacterial strain of qualification percentage ratio is the taxon belonging to test strains.The formulation of judgement criteria is on the basis of existing commercial two kinds of identification systems of generally acknowledging in the world, in conjunction with Chinese Shi Yuanxingchang bar section bacterial classification main biochemical type, serotype, genotype aspect carry out, being with Chinese characteristics property, is more suitable for the qualification result evaluation of domestic enterobacteriaceae bacterium than two kinds of identification systems of generally acknowledging.
Can all provide inconsistent biochemical test to the qualification of each test strains, basis for estimation judges with reference to the numerical value in probability matrix, and out single-row.
In result display, provide Chinese to taxon, Latin contrasts, and meets Chinese writing style.
1, biochemical test
First bacterium A to be measured is carried out gramstaining, oxydase, OF test, it is carried out to the qualification of Chang Gan section Cordycepps, chooses Gram-negative, other 23 kinds of biochemical tests that enterobacteriaceae bacterium that oxidase negative, OF are fermented type carries out except oxydase: phenylalanine deamination, wood sugar, raffinose, indoles, sucrose, ornithine decarboxylase, lactose, ONPG, urine enzyme, Citrate trianion, maltose, lysine decarboxylase, malonate, sorbyl alcohol, galactitol, pectinose, melibiose, rhamnosyl, Vitamin C2, MR, H
2s, trehalose, cellobiose.Record experimental result.
2, the calculating of percentage ratio is identified
After obtaining biochemical test result, calculate the qualification percentage ratio of 159 taxons and bacterium to be identified, owing to the positive probability of 159 taxons all cannot be listed as space is limited, so illustrate to be described with the tri-kinds of bacterium of Escherichiacoli, Citrobacterfreundii, the Enterobactercloacae in taxon and its biochemical test A, biochemical test B, biochemical test C.
Bacterium A biochemical test result to be measured and the biochemical positive probability results of Escherichiacoli, Citrobacterfreundii, Enterobactercloacae tri-kinds of bacterium as shown in table 1:
Table 1 biochemical test result and positive probability
| Bacterium name | Biochemical test A | Biochemical test B | Biochemical test C |
| Bacterium A to be measured | + | - | + |
| Escherichia coli | 98 | 1 | 90 |
| Citrobacter freundii | 33 | 78 | 0 |
| Enterobacter cloacae | 50 | 100 | 99 |
Biochemical test A, biochemical test B and biochemical test C belong to three kinds in above-mentioned 24 kinds of biochemical tests.
According to the biochemical test result of bacterium A to be measured, the result of three biochemical tests is respectively+,-,+, therefore according to foregoing method of calculation, the total frequency of occurrences=98 × (the 100-1) × 90=873180 of Escherichiacoli; The total frequency of occurrences=33 × (the 100-78) × 1=726 of Citrobacterfreundii; The total frequency of occurrences=50 × (100-100) × 99=50 × 1 × 99=4950 of Enterobactercloacae; The multinomial total frequency of occurrences=873180+726+4950=878856; Qualification percentage ratio=873180 ÷ 878856 × the 100=99.35 of Escherichiacoli; Qualification percentage ratio=726 ÷ 878856 × the 100=0.08 of Citrobacterfreundii; Qualification percentage ratio=4950 ÷ 878856 × the 100=0.56 of Enterobactercloacae.It is 24 because the taxon that the present invention includes is 159 and biochemical reaction, often identify that a strain bacterium needs all to carry out identifying to 159 taxons the calculating of percentage ratio, artificial computational effort is large, the progress of work will be subject to serious impact, therefore, computation process is carried out at corresponding software kit.
3 evaluation of result and qualification
In 159 taxons, according to the sequence of qualification percentage ratio size, list the first five taxon, judge according to evaluation of result standard.
As in this implementation column, be rearranged for according to qualification percentage ratio size: Escherichiacoli, Enterobactercloacae, Citrobacterfreundii.
Bacterium A qualification result to be measured is: colon bacillus (Escherichiacoli).
Inconsistent biochemical test is as shown in table 2:
The biochemical test that table 2 is inconsistent
| Escherichia coli | Citrobacter freundii | Enterobacter cloacae |
| + | Biochemical test A | Biochemical test B |
The checking of the numerical value rapid identification method degree of confidence of 4 food-borne pathogens Chang Gan section bacterial classifications
Experimental strain comprises 40 strain enterobacteriaceae reference cultures, and the bacterium source in actual sample is isolated Chang Gan section bacterial classification in nationwide is to the pollution survey of meat quail, fishery products, milk-product, vegetables, prepared food, quickfrozen food, the large based food of edible mushrooms, totally 7 and Pearl River water.These Chang Gan section bacterial classifications are identified according to the numerical value rapid identification method of enterobacteriaceae of the present invention, and with the MIDGNA of similar commercial prod France Mei Liai company API20E and Microgen company of Britain with or GNB, proved with the molecular biology PCR method of specific gene, 16SrDNA gene sequencing method.In 40 strain reference cultures and 1605 strain strain isolateds, the qualification result of each method as table 3,4,5,6:
Confirmable qualification result comprises fabulous, fine, good and acceptable qualification result, and uncertain qualification result comprises suspicious, unacceptable qualification result.Consistent qualification result and inconsistent qualification result are analyzed on confirmable result basis, if the present invention is consistent with results of comparison qualification result, be then consistent qualification result, vice versa.Preferably qualification result is that qualification result of the present invention is better than contrast; Poor qualification result is that result of the present invention is worse than contrast.Inconsistent qualification result is divided into the present invention and results of comparison to be the qualification result determined; Or result of the present invention is for determining, and contrast is not confirmable qualification result; Or result of the present invention can not be determined, and contrast is confirmable qualification result.All undesirable qualification result is then the present invention is all not confirmable qualification result with contrasting.
The numerical value rapid identification method of table 3 enterobacteriaceae of the present invention and MID method qualification result
| Result/strain number | The present invention (%) | MID(％) |
| Confirmable qualification result | 706(96.71) | 690(94.52) |
| Uncertain qualification result | 24(3.29) | 40(5.48) |
| Amount to | 730 | 730 |
The numerical value rapid identification method of table 4 enterobacteriaceae of the present invention and the difference of MIDGNA method qualification result
| Bacterial strain number | |
| Consistent qualification result | 678 |
| Inconsistent qualification result | 6 |
| The present invention is qualification result preferably | 28 |
| The qualification result that the present invention is poor | 11 |
| Neither desirable qualification result | 7 |
| Add up to | 730 |
The numerical value rapid identification method of table 5 enterobacteriaceae of the present invention and API20E method qualification result
| Result/strain number | The present invention | API 20E |
| Confirmable qualification result | 911(92.39％) | 863(87.53％) |
| Uncertain qualification result | 75(7.61％) | 123(12.47％) |
| Add up to | 986 | 986 |
The numerical value rapid identification method of table 6 enterobacteriaceae of the present invention and the difference of API20E method qualification result
| Bacterial strain number | |
| Consistent qualification result | 834 |
| Inconsistent qualification result | 17 |
| The present invention is qualification result preferably | 77 |
| The qualification result that the present invention is poor | 29 |
| Neither desirable qualification result | 29 |
| Add up to | 986 |
Both can well distinguish in the qualification without malonate citric acid fungus and kirschner citric acid fungus by the present invention, and the result of API20E can only be both results.Result is as shown in table 7:
The present invention can identify the level of biotype to the qualification of enterocolitis yersinia genus, and the present invention identifies biological 1A type to 128 strain enterocolitis yersinia genus, and API20E and MIDGNA+GNB can only identify the level of planting.
40 strain reference cultures, the present invention with contrast (API20E) good qualification result can be obtained.Carry out verifying with the molecular biology PCR method of specific gene for colon bacillus (544 strain), salmonella (308 strain) and E.sakazakii (154 strain) in experimental strain.Each bacterial strain PCR can amplify correct specific band, and the colon bacillus of the present invention's qualification, salmonella and E.sakazakii are respectively 527 strains, 308 strains, 154 strains; Colon bacillus, salmonella and E.sakazakii that contrast is identified are respectively 519 strains, 308 strains, 154 strains.The present invention with contrast the bacterial strain number no significant difference identified, but the present invention has 152 strains to identify subgenus-Salmonella enteritidis I in salmonella qualification, and API20E can only identify Salmonella, by serological identification, confirm that 152 strain Salmonellass are Salmonella enteritidis I really; The present invention has the qualification result qualification result as well of 47 strains when identifying Enterobacter sakazakii, but API20E needs increasing the qualification result that the basis of yellow pigment just can obtain, and therefore the present invention is better than contrasting API20E on the whole.
The checking of 16SrDNA gene sequencing method has been carried out to 270 Zhu Chang bar sections bacterium (except colon bacillus, salmonella and Enterobacter sakazakii).In conjunction with 16S qualification result, the present invention's strain isolated consistent with 16S result is 255 strains, has the result of 15 strains inconsistent; The consistent strain isolated of API20E and 16S qualification result is 210 strains, has the qualification result of 60 strains inconsistent; Wherein there is the result of 10 strain the present invention and API20E and 16S all inconsistent.Therefore from 16S qualification result, qualification result of the present invention is better than API20E.
Claims (3)
1. a numerical value rapid identification method for food-borne pathogens Chang Gan section bacterial classification, is characterized in that, comprise the following steps:
The probability that a, the kind determining Chang Gan section bacterium and at least 24 biochemical tests appearance thereof are positive, 24 described biochemical tests are: phenylalanine deamination, wood sugar, raffinose, indoles, sucrose, ornithine decarboxylase, lactose, ONPG, urine enzyme, Citrate trianion, maltose, lysine decarboxylase, malonate, sorbyl alcohol, galactitol, pectinose, melibiose, rhamnosyl, Vitamin C2, MR, H
2s, trehalose, cellobiose, oxydase;
24 biochemical tests in b, selection step a are as biochemical reaction required in numerical method:
C, set up the database of Chang Gan section bacterium, determine numerical method Computational Methods: according to the positive probability of the taxon selected and biochemical test, form a probability matrix;
Determine the method for calculation of total frequency of occurrences of each taxon, multinomial total occurrence frequency, qualification percentage ratio according to bacterium probability appraisal theory, specific formula for calculation is as follows:
Total frequency of occurrences of each taxon equals the product of the positive probability of each biochemical test corresponding to this taxon;
Multinomial total occurrence frequency equals the summation of total frequency of occurrences of each taxon;
Qualification percentage ratio equals the total frequency of occurrences of each taxon and is multiplied by 100 again divided by multinomial total occurrence frequency, and all qualification percentage ratio sums should equal 100;
After d, test strains pure culture, carry out gramstaining, oxydase, OF tests, and select gramstaining be feminine gender, oxidase negative, OF is 24 kinds of biochemical tests that the bacterial strain of fermented type carries out step a;
After 24 kinds of biochemical test results of e, acquisition test strains, according to yin, yang, the unknown-value of biochemical reaction, according to method of calculation, taxons all in database are calculated, first biochemical test value is converted to probable value, i.e. the positive probability value of experimental result to be the positive be exactly then corresponding biochemical test; If feminine gender is the 100 positive probability values deducting biochemical test then, if the above results 0 or 100, then replace by approximation 1 or 99, again according to the method for calculation of step c calculate each taxon total frequency of occurrences, multinomial total occurrence frequency, identify that if percentage ratio a certain biochemical test result is for unknown or smudgy, then this biochemical test item does not participate in calculating;
F, according to calculation result, to judge identification of strains result to be measured: the size according to qualification percentage ratio sorts, the highest corresponding bacterial strain of qualification percentage ratio is the taxon belonging to test strains.
2. the numerical value rapid identification method of food-borne pathogens Chang Gan section according to claim 1 bacterial classification, is characterized in that, all provides inconsistent biochemical test and out single-row to the qualification of test strains.
3. the numerical value rapid identification method of food-borne pathogens Chang Gan section according to claim 1 bacterial classification, is characterized in that, in result display, provide Chinese to taxon, Latin contrasts.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510866404.3A CN105483207B (en) | 2015-11-30 | 2015-11-30 | Method for rapidly identifying value of enterobacter strains of food-borne pathogenic bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510866404.3A CN105483207B (en) | 2015-11-30 | 2015-11-30 | Method for rapidly identifying value of enterobacter strains of food-borne pathogenic bacteria |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105483207A true CN105483207A (en) | 2016-04-13 |
| CN105483207B CN105483207B (en) | 2020-03-27 |
Family
ID=55670478
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510866404.3A Active CN105483207B (en) | 2015-11-30 | 2015-11-30 | Method for rapidly identifying value of enterobacter strains of food-borne pathogenic bacteria |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105483207B (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1081717A (en) * | 1993-06-14 | 1994-02-09 | 何林 | Biochemical identification carton for bacteria |
| WO1996040861A1 (en) * | 1995-06-07 | 1996-12-19 | Biolog, Inc. | Microbiological media for isolation and identification of enteric pathogens such as e. coli and salmonella |
| CN1186858A (en) * | 1997-01-02 | 1998-07-08 | 许仁爱 | Identifying method of bacteria and fungus and its test card |
| CN1268668A (en) * | 1999-03-29 | 2000-10-04 | 许仁爱 | Blank card for microbial detection and its microbial detection test culture medium card |
| CN1884482A (en) * | 2006-06-02 | 2006-12-27 | 华中师范大学 | Aerogenic enteric bacilli secreting alga-dissolving substance and its uses in myxophyceae algal tufa |
| CN101487039A (en) * | 2009-01-23 | 2009-07-22 | 山东出入境检验检疫局检验检疫技术中心 | Bacteria screening and identifying method |
| CN102206604A (en) * | 2011-04-28 | 2011-10-05 | 南京大学 | Enterobacter agglomerans strain and application thereof in weeding |
-
2015
- 2015-11-30 CN CN201510866404.3A patent/CN105483207B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1081717A (en) * | 1993-06-14 | 1994-02-09 | 何林 | Biochemical identification carton for bacteria |
| WO1996040861A1 (en) * | 1995-06-07 | 1996-12-19 | Biolog, Inc. | Microbiological media for isolation and identification of enteric pathogens such as e. coli and salmonella |
| CN1186858A (en) * | 1997-01-02 | 1998-07-08 | 许仁爱 | Identifying method of bacteria and fungus and its test card |
| CN1268668A (en) * | 1999-03-29 | 2000-10-04 | 许仁爱 | Blank card for microbial detection and its microbial detection test culture medium card |
| CN1884482A (en) * | 2006-06-02 | 2006-12-27 | 华中师范大学 | Aerogenic enteric bacilli secreting alga-dissolving substance and its uses in myxophyceae algal tufa |
| CN101487039A (en) * | 2009-01-23 | 2009-07-22 | 山东出入境检验检疫局检验检疫技术中心 | Bacteria screening and identifying method |
| CN102206604A (en) * | 2011-04-28 | 2011-10-05 | 南京大学 | Enterobacter agglomerans strain and application thereof in weeding |
Non-Patent Citations (3)
| Title |
|---|
| 周用直 等主编: "《感染性腹泻病原检测技术 》", 31 August 1996, 郑州：中原农民出版社 * |
| 温建新等: "家兔致病性大肠杆菌的分离鉴定与药敏试验", 《中国畜牧兽医》 * |
| 苏丽春等: "一种新的细菌生化鉴定系统Microgen ID", 《中国卫生检验杂志》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105483207B (en) | 2020-03-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Janda et al. | The changing face of the family Enterobacteriaceae (Order:“Enterobacterales”): New members, taxonomic issues, geographic expansion, and new diseases and disease syndromes | |
| Weinroth et al. | Considerations and best practices in animal science 16S ribosomal RNA gene sequencing microbiome studies | |
| Farmer 3rd et al. | Escherichia fergusonii and Enterobacter taylorae, two new species of Enterobacteriaceae isolated from clinical specimens | |
| Morales-López et al. | Enterobacteria in the 21st century: a review focused on taxonomic changes | |
| Bjerke et al. | Exploring the brine microbiota of a traditional Norwegian fermented fish product (Rakfisk) from six different producers during two consecutive seasonal productions | |
| Hascoët et al. | Microbial ecology evaluation of an iberian pig processing plant through implementing SCH sensors and the influence of the resident microbiota on Listeria monocytogenes | |
| Asselin et al. | Rahnella spp. are commonly isolated from onion (Allium cepa) bulbs and are weakly pathogenic | |
| Callicott et al. | Broiler Campylobacter contamination and human campylobacteriosis in Iceland | |
| Habib et al. | Performance characteristics and estimation of measurement uncertainty of three plating procedures for Campylobacter enumeration in chicken meat | |
| CN105483207A (en) | Rapid numerical identification method for enterobacteria of food-borne pathogenic bacteria | |
| Budiarso et al. | Detection of coliforms and enteric pathogens in favorite snack food sold in Yogyakarta City | |
| Line et al. | Comparison of an automated most-probable-number technique with traditional plating methods for estimating populations of total aerobes, coliforms, and Escherichia coli associated with freshly processed broiler chickens | |
| CN109187385A (en) | A kind of method of high light spectrum image-forming identification microorganism | |
| Amakelew et al. | Microbial quality of cow milk in Dawa Chefa district, Amhara Region, Ethiopia | |
| Tsai et al. | Development of Shortened Enrichment Methods for Detection of Salmonella Typhimurium Spiked in Milk | |
| Mainello-Land et al. | Multilocus sequence and phenotypic analysis of Pectobacterium and Dickeya type strains for identification of soft rot Pectobacteriaceae from symptomatic potato stems and tubers in Pennsylvania | |
| CN113512601B (en) | Molecular targets for screening for Proteus and quantitative detection methods | |
| Cole | Databases in modern food microbiology | |
| WO2019091497A1 (en) | Earthworm juvenile individual and group quality evaluation methods | |
| Al-Nasrawy et al. | Molecular Research Comparing the Probabilities of Burkholderia Cepacia Bacterium Diagnosis Procedures. | |
| CN105861621A (en) | Method for identifying bacterium numerical value of common Listeria monocytogenes in food | |
| Arunachalam et al. | Molecular phylogenetic approach for classification of Salmonella typhi | |
| Buller et al. | Identification of bacteria from aquatic animals | |
| FUNG et al. | Evaluation of Candida isolation medium in food hygiene in Hungary | |
| Wibowo et al. | Identification of Pathogenic Bacteria on Spiny Lobster (Panulirus homarus) at Quality Control and Fishery Product Safety Agency (BKIPM) of Bengkulu |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP03 | Change of name, title or address |
Address after: Guangzhou City, Guangdong province 510070 martyrs Road No. 100 building No. 56 Co-patentee after: GUANGDONG HUANKAI BIOTECHNOLOGY Co.,Ltd. Patentee after: GUANGDONG INSTITUTE OF MICROBIOLOGY (GUANGDONG DETECTION CENTER OF MICROBIOLOGY) Co-patentee after: GUANGDONG HUANKAI MICROBIAL SCI & TECH. Co.,Ltd. Address before: Guangzhou City, Guangdong province 510070 martyrs Road No. 100 Co-patentee before: GUANGDONG HUANKAI BIOTECHNOLOGY Co.,Ltd. Patentee before: GUANGDONG INSTITUTE OF MICROBIOLOGY Co-patentee before: GUANGDONG HUANKAI MICROBIAL SCI & TECH. Co.,Ltd. |
|
| CP03 | Change of name, title or address |