CN101724682A - Staining fluid and method for rapidly detecting novel cryptococcus - Google Patents
Staining fluid and method for rapidly detecting novel cryptococcus Download PDFInfo
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- CN101724682A CN101724682A CN200910154521A CN200910154521A CN101724682A CN 101724682 A CN101724682 A CN 101724682A CN 200910154521 A CN200910154521 A CN 200910154521A CN 200910154521 A CN200910154521 A CN 200910154521A CN 101724682 A CN101724682 A CN 101724682A
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Abstract
The invention relates to staining fluid and a method for rapidly detecting novel Cryptococcus. The staining fluid comprises 0.5-3.0g of AZO-blue, 1.0-10.0g of glacial acetic acid and 90-110g of distilled water. The method for rapidly detecting novel cryptococcus comprises the steps of preparation of the staining fluid, mixing, detection, counting and the like. The invention has the advantages of simple, convenient, rapid, sensitive and accurate detection, high specificity, easy preservation of the staining fluid, short staining time and excellent staining effect.
Description
Technical field
The invention belongs to the detection range of a kind of Cryptococcus neoformans, staining fluid of especially a kind of rapid detection Cryptococcus neoformans and method.
Background technology
Diagnosis for Cryptococcus neoformans mainly depends on prepared Chinese ink smear and fungus culture at present, with detecting to making a definite diagnosis foundation of pathogenic agent.Aspect quick diagnosis, though the direct smear ink dyeing is easy, mirror is observed difficulty relatively down, thereby positive rate is low, even foreign literature reports that positive rate also has only 53%～56% under continuous several times inspection situation, there is certain false positive simultaneously, and is difficult to carry out the cryptococcus counting.The tradition fungus culture generally needs a couple of days, can reach more than 90% though cultivate positive rate, because of long delay diagnosis of cycle.Animal inoculation pvaccination can detect that the clinical height of part is suspected and smear, the equal negative patients of cultivation, and this method is consuming time longer, and is very little for the quick diagnosis meaning, clinically seldom uses.In recent years, along with the fast development of Protocols in Molecular Biology and immunological technique is that new approach has been opened up in the diagnosis of Cryptococcus neoformans, report all confirms to utilize immunological technique and round pcr to have higher accuracy and susceptibility aspect quick diagnosis both at home and abroad, but because the test kit difference that adopts, the result is often variant, certain false positive and false negative rate are arranged, and because immunological technique and round pcr detect Cryptococcus neoformans cost height, operation is than dyeing microscopic examination complexity, do not belong to the gold standard of diagnosing Cryptococcus neoformans again, also be unfavorable in the small towns and basic hospital is carried out, thereby in the clinical diagnosis work of clinical laboratory, be not used widely.
Summary of the invention
The present invention is directed to the shortcoming of above-mentioned prior art, provide a kind of quick and precisely, staining fluid and the method for the rapid detection Cryptococcus neoformans of high specific.
The present invention solves the technical scheme that its technical problem adopts:
The staining fluid of this rapid detection Cryptococcus neoformans comprises, 0.5-3.0 gram AZO-blue, 1.0-10.0 gram Glacial acetic acid, 90-110 restrain distilled water.
The method of this rapid detection Cryptococcus neoformans may further comprise the steps,
1) makes staining fluid: utilize 0.5-3.0 gram AZO-blue, 1.0-10.0 gram Glacial acetic acid, 90-110 gram distilled water to make staining fluid;
2) mix: sample preparations to be checked is become behind the bacteria suspension and above-mentioned staining fluid mixing;
3) detect: above-mentioned mixing liquid is carried out microscopy.
Described sample to be checked can be tissue homogenate or bacterium colony.
The method of another kind of rapid detection Cryptococcus neoformans may further comprise the steps,
1) makes staining fluid: utilize 0.5-3.0 gram AZO-blue, 1.0-10.0 gram Glacial acetic acid, 90-110 gram distilled water to make staining fluid;
2) mix: with behind the specimen centrifuge to be checked with above-mentioned staining fluid mixing;
3) counting: above-mentioned mixing liquid is added tally accurately count.
Described sample to be checked can be cerebrospinal fluid or ascites pleural fluid.
The method of another kind of rapid detection Cryptococcus neoformans may further comprise the steps,
1) makes staining fluid: utilize 0.5-3.0 gram AZO-blue, 1.0-10.0 gram Glacial acetic acid, 90-110 gram distilled water to make staining fluid;
2) mix: sample disposal to be checked and above-mentioned staining fluid mixing;
3) detect: above-mentioned mixing liquid is carried out microscopy.
4) counting: as detect the existence of Cryptococcus neoformans, can more above-mentioned mixed solution be added tally and accurately count.
The effect that the present invention is useful is: one, detect easy quick, sensitive and accurate, have a high specific; Two, staining fluid is easily preserved; Three, dyeing time is short, and Color is excellent.
Description of drawings
Fig. 1 is the rapid dye liquor Color figure (10 * 100) of Cryptococcus neoformans;
Fig. 2 is coexist rapid dye liquor Color figure (10 * 40) in the counting cell of Cryptococcus neoformans, Candida albicans, white corpuscle;
Fig. 3 is the ink dyeing design sketch (10 * 100) of Cryptococcus neoformans;
Fig. 4 is that Cryptococcus neoformans is used the rapid dye liquor Color figure (10 * 40) that configured a year;
Fig. 5 is that Cryptococcus neoformans is used the configuration rapid dye liquor Color figure (10 * 40) of what a month;
Fig. 6 is the rapid dye liquor Color figure (10 * 40) of Cryptococcus neoformans with fresh configuration;
Fig. 7 is the rapid dye liquor 1min Color figure (10 * 40) of Cryptococcus neoformans;
Fig. 8 is the rapid dye liquor 5min Color figure (10 * 40) of Cryptococcus neoformans;
Fig. 9 is the rapid dye liquor 30min Color figure (10 * 40) of Cryptococcus neoformans;
Embodiment
The invention will be further described below in conjunction with accompanying drawing:
The staining fluid of a kind of rapid detection Cryptococcus neoformans,
Embodiment 1: by the preparation of following weight proportion raw material, and AZO-blue 1 gram, Glacial acetic acid 2 grams, distilled water 97 grams.
Embodiment 2: by the preparation of following weight proportion raw material, and AZO-blue 2 grams, Glacial acetic acid 5 grams, distilled water 93 grams.
Embodiment 3: by the preparation of following weight proportion raw material, and AZO-blue 0.5 gram, Glacial acetic acid 1 gram, distilled water 99 grams.
Embodiment 4: by the preparation of following weight proportion raw material, and AZO-blue 2.5 grams, Glacial acetic acid 10 grams, distilled water 110 grams.
The dyeing process of a kind of rapid detection Cryptococcus neoformans,
Embodiment 1: its step is,
1) makes staining fluid: utilize above-mentioned any one proportioning of staining fluid embodiment to make staining fluid;
2) mix: sample preparations such as tissue homogenate or bacterium colony are become behind the bacteria suspension and above-mentioned staining fluid mixing;
3) detect: above-mentioned mixing liquid is carried out microscopy, to determine existing of Cryptococcus neoformans.
As Fig. 1, visible Cryptococcus neoformans is dyed blueness or mazarine, and its outside pod membrane does not dye, and becomes the ring-type of light.
Embodiment 2: its step is,
1) makes staining fluid: utilize the proportioning of above-mentioned any one embodiment to make staining fluid;
2) mix: with behind the specimen centrifuges such as cerebrospinal fluid, ascites pleural fluid with above-mentioned staining fluid mixing;
3) counting: above-mentioned mixing liquid is added tally accurately count, to determine the quantity of Cryptococcus neoformans.
As Fig. 2, as seen Candida albicans is white in color in tally, and the big look of white corpuscle body is shallow, and white corpuscle and other common fungies are not dyeed, and is easy to Cryptococcus neoformans and white corpuscle and other common fungi differentiations.
Embodiment 3: its step is,
1) makes staining fluid: utilize the proportioning of above-mentioned any one embodiment to make staining fluid;
2) mix: with behind the specimen centrifuges such as cerebrospinal fluid, ascites pleural fluid with above-mentioned staining fluid mixing;
3) detect: above-mentioned mixing liquid is added mixing liquid carry out microscopy.
4) counting:, can more above-mentioned mixing liquid be added tally and accurately count as having detected Cryptococcus neoformans by above-mentioned detection.
The Color contrast of staining fluid of the present invention and other staining agents:
As Fig. 3, Cryptococcus neoformans is under the dyeing of prepared Chinese ink, and pod membrane is bright, but not as dark with the painted coating of the present invention, internal structure is also painted clear not as the present invention.
Staining fluid stability assessment of the present invention:
As Fig. 4-6, visible configuration 1 year, configuration one month, freshly prepared staining fluid all have good Color and specificity to Cryptococcus neoformans.
Color stability assessment of the present invention:
As Fig. 7-9, in the visible sample dyeing 30min good stability is arranged, along with dyeing time prolongs, the painted meeting of Cryptococcus neoformans is deepened gradually, and good Color and specificity are arranged.
In addition to the implementation, the present invention can also have other embodiments.All employings are equal to the technical scheme of replacement or equivalent transformation formation, all drop on the protection domain of requirement of the present invention.
Claims (6)
1. the staining fluid of a rapid detection Cryptococcus neoformans is characterized in that: described staining fluid comprises that 0.5-3.0 gram AZO-blue, 1.0-10.0 gram Glacial acetic acid, 90-110 restrain distilled water.
2. the method for a rapid detection Cryptococcus neoformans is characterized in that: said method comprising the steps of,
1) makes staining fluid: utilize 0.5-3.0 gram AZO-blue, 1.0-10.0 gram Glacial acetic acid, 90-110 gram distilled water to make staining fluid;
2) mix: sample preparations to be checked is become behind the bacteria suspension and above-mentioned staining fluid mixing;
3) detect: above-mentioned mixing liquid is carried out microscopy.
3. the method for rapid detection Cryptococcus neoformans according to claim 2 is characterized in that: described sample to be checked is tissue homogenate or bacterium colony.
4. the method for a rapid detection Cryptococcus neoformans is characterized in that: said method comprising the steps of,
1) makes staining fluid: utilize 0.5-3.0 gram AZO-blue, 1.0-10.0 gram Glacial acetic acid, 90-110 gram distilled water to make staining fluid;
2) mix: with behind the specimen centrifuge to be checked with above-mentioned staining fluid mixing;
3) counting: above-mentioned mixing liquid is added tally accurately count.
5. the method for rapid detection Cryptococcus neoformans according to claim 4 is characterized in that: described sample to be checked is cerebrospinal fluid or ascites pleural fluid.
6. the method for a rapid detection Cryptococcus neoformans is characterized in that: said method comprising the steps of,
1) makes staining fluid: utilize 0.5-3.0 gram AZO-blue, 1.0-10.0 gram Glacial acetic acid, 90-110 gram distilled water to make staining fluid;
2) mix: sample disposal to be checked and above-mentioned staining fluid mixing;
3) detect: above-mentioned mixing liquid is carried out microscopy;
4) counting: as detect the existence of Cryptococcus neoformans, more above-mentioned mixed solution is added tally and accurately count.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2009101545211A CN101724682B (en) | 2009-11-10 | 2009-11-10 | Staining fluid and method for rapidly detecting novel cryptococcus |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2009101545211A CN101724682B (en) | 2009-11-10 | 2009-11-10 | Staining fluid and method for rapidly detecting novel cryptococcus |
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| CN101724682A true CN101724682A (en) | 2010-06-09 |
| CN101724682B CN101724682B (en) | 2012-06-06 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102816705A (en) * | 2012-05-16 | 2012-12-12 | 康颖倩 | Saccharomycete of basidiomycete and cultural method thereof |
| CN107167357A (en) * | 2017-06-22 | 2017-09-15 | 崔舜� | Deep fungal morphology quick determination method and reagent based on counterstain |
| CN108676838A (en) * | 2018-04-11 | 2018-10-19 | 马爽 | A kind of cryptococcus capsule stain liquid and its preparation and application |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1039826C (en) * | 1994-01-25 | 1998-09-16 | 中国医学科学院皮肤病研究所 | Appraising method for neogenesis cryptococcus and used culture-medium thereof |
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2009
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102816705A (en) * | 2012-05-16 | 2012-12-12 | 康颖倩 | Saccharomycete of basidiomycete and cultural method thereof |
| CN107167357A (en) * | 2017-06-22 | 2017-09-15 | 崔舜� | Deep fungal morphology quick determination method and reagent based on counterstain |
| CN107167357B (en) * | 2017-06-22 | 2020-05-15 | 崔舜� | Deep fungus morphology rapid detection method and reagent based on contrast dyeing |
| CN108676838A (en) * | 2018-04-11 | 2018-10-19 | 马爽 | A kind of cryptococcus capsule stain liquid and its preparation and application |
| CN108676838B (en) * | 2018-04-11 | 2021-09-24 | 马爽 | Cryptococcus capsular staining solution and preparation and use methods thereof |
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| CN101724682B (en) | 2012-06-06 |
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