CN105713955A - Culture medium for identifying group-B streptococcus - Google Patents
Culture medium for identifying group-B streptococcus Download PDFInfo
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- CN105713955A CN105713955A CN201610178973.3A CN201610178973A CN105713955A CN 105713955 A CN105713955 A CN 105713955A CN 201610178973 A CN201610178973 A CN 201610178973A CN 105713955 A CN105713955 A CN 105713955A
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- 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
- C12Q1/14—Streptococcus; Staphylococcus
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- 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
- C12Q1/045—Culture media therefor
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Abstract
The invention discloses a culture medium for identifying group-B streptococcus. The culture medium in per 100 ML water comprises, by mass, 3 g of proteose peptone, 0.2 g of KH2PO4, 0.4 g of Na2HPO4, 4 g of crude rice flour, 1.3 g of agar, 0.2 g of antibiotic, 0.2 g of brain-heart baptist extraction liquid medium and 2 g of horse sera. According to the culture medium, scribing detection can be directly carried out through a sampled to-be-detected swab; compared with the condition that pure bacterial strains need to be identified in the prior art, the detection accuracy and the detection efficiency of the group-B streptococcus are improved through the culture medium; An application method is convenient to operate, and special instruments and devices are not required; the culture medium is suitable for hospital and particularly suitable for applications of basic medical institutions, sampling work of samples is convenient, and meanwhile the detection effect of the group-B streptococcus is guaranteed.
Description
Technical field
The present invention relates to a kind of microbiological culture media, particularly relate to a kind of B race streptococcus selective medium.
Background technology
B race streptococcus (GBS), also known as B group streptococcus or streptococcus agalactiae, belong to gram positive bacteria, it is perch the normal flora at human gastrointestinal tract and Lower genital tract, since 20 century 70s, GBS is increasingly becoming one of important pathogenic bacteria of neonate serious infections, has caused global extensive concern.It is the main cause causing the severe infections such as fetus at perinatal stage intrauterine infection, pneumonia of newborn, septicemia, meningitis that GBS infects.Premature labor can be caused, the serious consequence such as fetal development slow (low birth weight infant), premature rupture of fetal membrane and late abortion, in perinatal stage medical science, account for critical role.
At present, mainly having two kinds of methods whether pregnant and lying-in women are carried in GBS carries out examination, the first is to use detection kit to carry out quick diagnosis.But it is all higher to requirement and the cost of operation to carry out quick diagnosis by detection kit, limits the popularization and application of detection kit.The second, namely current modal method, be utilize culture medium that the sample obtained is cultivated, and then passes through direct observational method, Gram staining method, catalase test, collaborative haemolysis (CAMP) test etc. judge whether to infect GBS.But anemia of pregnant woman's reproductive tract and rectum have various bacteria to exist, and current culture medium can not effectively suppress the growth of other bacterium beyond GBS, these bacteria growths are fast, often make GBS bacteria growing be suppressed, cause GBS bacterium missing inspection.For this, Clinical detection uses existing B race streptococcus differential medium carry out B race streptococcus when identifying, it is necessary to the first bacterial strain purification enrichment culture to sampling, then just can identify, such program is loaded down with trivial details, and loses time and manpower.The B race streptococcus examination culture medium of the Mei Liai company of current Clinical practice, what it mainly adopted is the method for substrate colour developing, relatively costly, price.
Summary of the invention
In order to overcome prior art Problems existing, the invention provides one can the streptococcic culture medium of Rapid identification B race.
The technical solution adopted in the present invention is:
A kind of B race streptococcus differential medium, every 100ML water includes the composition of following quality: proteose peptone 3g, KH2PO40.2g, Na2HPO40.4g, rough rice flour 4g, agar 1.3g, antibiotic 0.2g, the brain-heart leaching extraction liquid culture medium 0.2g, horse serum 2g.
Further, described antibiotic includes gentamycin, nalidixic acid and amphotericin B.
Further, described antibiotic in the medium content be respectively as follows: gentamycin: 8 × 10-4G/ml, nalidixic acid: 10 × 10-4G/ml, amphotericin B: 2 × 10-4g/ml。
Further, described rough rice flour is for making after being pulverized by brown rice.
Further, the preparation method of described culture medium comprises the following steps: (1) is by the rough rice flour of formula ratio, proteose peptone, KH2PO4、Na2HPO4, agar and brain-heart leaching extraction liquid culture medium be dissolved in that 100mL is mono-steams water, correct PH to 7.2 ± 0.2 with 10mol/L sodium hydroxide；(2) autoclave sterilization, to be cooled to about 60 DEG C time move in Biohazard Safety Equipment；(3) add antibiotic and horse serum shakes up, then aforesaid liquid culture medium is poured in sterilized petri dishes, pack after solidification, in 4 DEG C of preservations.
The invention have the benefit that
The B race streptococcus differential medium of the present invention uses rough rice flour but not commercially available deep processing rice flour.Research finds, the streptococcus colour developing of rough rice flour Cai Nengshi B race.The culture medium of the present invention can utilize the swab to be checked of sampling to be made directly line detection, purebred bacterial strain need to be identified by hinge structure, the culture medium of the present invention improves detection accuracy and the detection efficiency of B group streptococcus, and using method of the present invention is simple to operate, it is not necessary to special instrument and equipment.Being adapted at hospital, especially basic medical unit application, facilitates the sampling work of sample, ensure that the detection effect of B group streptococcus simultaneously.
In order to be more fully understood that and implement, the following detailed description of the present invention.
Detailed description of the invention
The preparation of [embodiment 1] B race streptococcus differential medium
B race streptococcus differential medium in the present embodiment, every 100ML water includes the composition of following quality: proteose peptone 3g, KH2PO40.2g, Na2HPO40.4g, rough rice flour 4g, agar 1.3g, antibiotic 0.2g, the brain-heart leaching extraction liquid culture medium 0.2g, horse serum 2g.
Its compound method is: by the rough rice flour of formula ratio, proteose peptone, KH in conical flask2PO4、Na2HPO4, agar and brain-heart leaching extraction liquid culture medium be dissolved in that 100mL is mono-steams water, correct PH to 7.2 ± 0.2 with 10mol/L sodium hydroxide；.Bandage and put 121 DEG C of pressure cooker sterilizing 20min, to be cooled to about 60 DEG C time conical flask is moved in Biohazard Safety Equipment.Adding antibiotic and horse serum shakes up, then aforesaid liquid culture medium be poured in the sterilized petri dishes of internal diameter 9cm, culture medium height is about 0.5cm.Packaging after to be solidified, in 4 DEG C of preservations.
The application of [embodiment 2] B race streptococcus differential medium
Test procedure: swab to be checked adopts streaking inoculation to inoculate this culture medium.Directly rule with swab in firstth district, second and third, four district's inoculating loops rule；Cultivate 18h～24h observed result for 37 DEG C.
Result judges: typical case B race streptococcus as orange colonies, has colonies typical can report detection B race streptococcus on flat board during detection.
The streptococcus Rapid identification culture medium result critical table of B race
[embodiment 3] B race streptococcus differential medium inhibitory action to other mushrooms
The culture medium flat plate of the present invention is inoculated various types of antibacterial and fungus, observes its growing state as follows
| Strain name | Growing state | Strain name | Growing state |
| Staphylococcus aureus | - | Salmonella typhi | - |
| Alpha streptococcus | - | Dysentery bacterium D15 | - |
| γ-streptococcus | + | Escherichia coli | - |
| Bacillus pyocyaneus | - | Corynebacterium belfantii | - |
| Clostridium perfringen | - | Staphylococcus epidermidis | - |
| Bacillus proteus | - | Salmonella paratyphi B | - |
| Streptococcus pneumoniae | - | Candida albicans | - |
Strain do not grow use (-) represent；Strain have growth with (+) represent
Wherein, γ-streptococcus grows, and bacterium colony whitening color does not affect observed result.
[embodiment 4] clinical trial
19-65 year women 183 example is randomly choosed in the attached First Hospital of University Of Nanhua, the secretions specimen of reproductive tract is gathered with common cotton swab, the B race streptococcus differential medium of the present invention and the B race streptococcus examination culture medium of Mei Liai (Shanghai) biological product company limited production is used to detect respectively, for result difference group, re-use third party's reagent and detect.
Testing result, recall rate 19.13% (35/183) of the present invention, contrast agent verification and measurement ratio is the B race streptococcus differential medium recall rate 92% of 21.88% (40/183), false positive rate 0%.
The invention is not limited in above-mentioned embodiment, if to the various changes of the present invention or deformation without departing from the spirit and scope of the present invention, if these are changed and deform within the claim and the equivalent technologies scope that belong to the present invention, then the present invention is also intended to comprise these changes and deformation.
Claims (5)
1. a B race streptococcus differential medium, it is characterised in that every 100ML water includes the composition of following quality: proteose peptone 3g, KH2PO40.2g, Na2HPO40.4g, rough rice flour 4g, agar 1.3g, antibiotic 0.2g, the brain-heart leaching extraction liquid culture medium 0.2g, horse serum 2g.
2. B race according to claim 1 streptococcus differential medium, it is characterised in that described antibiotic includes gentamycin, nalidixic acid and amphotericin B.
3. described in claim 2 B race streptococcus differential medium, it is characterised in that described antibiotic in the medium content be respectively as follows: gentamycin: 8 × 10-4G/ml, nalidixic acid: 10 × 10-4G/ml, amphotericin B: 2 × 10-4g/ml。
4. B race according to claim 1 streptococcus differential medium, it is characterised in that described rough rice flour is for making after being pulverized by brown rice.
5. the preparation method of the B race streptococcus differential medium described in claim 1, comprises the following steps: (1) is by the rough rice flour of formula ratio, proteose peptone, KH2PO4、Na2HPO4, agar and brain-heart leaching extraction liquid culture medium be dissolved in that 100mL is mono-steams water, correct PH to 7.2 ± 0.2 with 10mol/L sodium hydroxide；(2) autoclave sterilization, to be cooled to about 60 DEG C time move in Biohazard Safety Equipment；(3) add antibiotic and horse serum shakes up, then aforesaid liquid culture medium is poured in sterilized petri dishes, pack after solidification, in 4 DEG C of preservations.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610178973.3A CN105713955B (en) | 2016-03-25 | 2016-03-25 | A kind of B races streptococcus differential medium |
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| CN201610178973.3A CN105713955B (en) | 2016-03-25 | 2016-03-25 | A kind of B races streptococcus differential medium |
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| CN105713955A true CN105713955A (en) | 2016-06-29 |
| CN105713955B CN105713955B (en) | 2017-06-06 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434843A (en) * | 2016-10-27 | 2017-02-22 | 江南大学 | Culture medium for selectively separating and culturing group B streptococci and preparation method thereof |
| CN111423997A (en) * | 2020-03-31 | 2020-07-17 | 广州市妇女儿童医疗中心（广州市妇幼保健院、广州市儿童医院、广州市妇婴医院、广州市妇幼保健计划生育服务中心） | Group B streptococcus enriched broth and application thereof |
| CN114645072A (en) * | 2022-05-18 | 2022-06-21 | 江苏达伯药业有限公司 | Liquid group B streptococcus selective chromogenic medium and preparation method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104388527A (en) * | 2014-11-24 | 2015-03-04 | 苏州嘉禧萝生物科技有限公司 | Streptococcus agalactiae selective chromogenic culture medium and detection test paper thereof |
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2016
- 2016-03-25 CN CN201610178973.3A patent/CN105713955B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104388527A (en) * | 2014-11-24 | 2015-03-04 | 苏州嘉禧萝生物科技有限公司 | Streptococcus agalactiae selective chromogenic culture medium and detection test paper thereof |
Non-Patent Citations (2)
| Title |
|---|
| MANUEL DE LA ROSA 等: ""New Granada Medium for Detection and Identification of Group B Streptococci"", 《JOURNAL OF CLINICAL MICROBIOLOGY》 * |
| MANUEL DE LA ROSA等: ""Granada Medium for Detection and Identification of Group B Streptococci"", 《JOURNAL OF CLINICAL MICROBIOLOGY》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434843A (en) * | 2016-10-27 | 2017-02-22 | 江南大学 | Culture medium for selectively separating and culturing group B streptococci and preparation method thereof |
| CN111423997A (en) * | 2020-03-31 | 2020-07-17 | 广州市妇女儿童医疗中心（广州市妇幼保健院、广州市儿童医院、广州市妇婴医院、广州市妇幼保健计划生育服务中心） | Group B streptococcus enriched broth and application thereof |
| CN111423997B (en) * | 2020-03-31 | 2022-02-11 | 广州市妇女儿童医疗中心（广州市妇幼保健院、广州市儿童医院、广州市妇婴医院、广州市妇幼保健计划生育服务中心） | Group B streptococcus enriched broth and application thereof |
| CN114645072A (en) * | 2022-05-18 | 2022-06-21 | 江苏达伯药业有限公司 | Liquid group B streptococcus selective chromogenic medium and preparation method thereof |
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