CN101845478A - Neutralization isotonic bacteria and virus mouth wash sampling liquid - Google Patents

Abstract

The invention relates to neutralization isotonic bacteria and virus mouth wash sampling liquid, in particular to a formula and a preparation method of neutral and isotonic mouth wash sampling liquid for treating hand-foot-and-mouth disease and respiratory pathogenic bacteria and viruses. The neutralization isotonic bacteria and virus mouth wash sampling liquid has the following advantages that: (1) the formula is simple, the preparation is easy, and qualified products can be prepared in a basic laboratory; (2) clinical application effects are good, and the sampling problem that discomfort such as vomiting is easily caused when a throat swab is used for sampling can be solved; (3) isotonic solution of sugar and salt is harmless to human body; and (4) nutrients are rich, residual antibiotic in a sample can be prevented, and the aim of improving the positive rate of virus isolation in a laboratory is achieved.

Description

In and isotonic bacteria and virus mouth wash sampling liquid

Technical field the present invention relates to a kind of in and isotonic bacteria and virus mouth wash sampling liquid; Particularly a kind ofly be used for prescription and the preparation method that the sample solution of gargling is oozed in hand foot mouth disease and respiratory tract pathogenic bacterium, virus neutralization etc.Hand foot mouth disease has another name called dermexanthesis bubble stomatitis, it is a kind of children's transmissible disease that China occurs in recent years, cause by multiple enterovirus infection, with brothers, buttocks and front portion, oral cavity is the eruption disease of predilection site, can betide the four seasons, summer and autumn is easily popular, and this disease is a disease of viral infection, prognosis bona behind antiviral and symptomatic treatment seldom has complication.But the patient with severe symptoms can merge myocarditis and meningitis threat to life.It was reported that total case load that hand foot mouth disease takes place Taiwan in 1998 has reached 12.9 ten thousand, severe 405 examples wherein, dead 78 examples; 91% age＜5 year old in the critical case, 70% merges viral encephalitis.Throat swab HANKS liquid is adopted in this sick sample examination more, its advantage is as keeping liquid, enterovirus that can fine preservation Picornaviridae (for example: 4,5,7,9,10,16 types of Coxsackie virus (Coxsackie virus) A group, 2,5 types of Coxsackie B virus group; Part Echo virus (ECHO-viruses) and enterovirns type 71).Improve the isolating positive rate of zoo virus.For liquid is preserved in first sampling of recommending in Ministry of Health's hand foot mouth disease sampling standard.Its shortcoming is the complexity of filling a prescription, the cost height; When adopting the throat swab sampling, cause child's vomiting reflex easily, sense of discomfort etc.As collutory, can be swallowed down by the child, cause untoward reaction (containing magnesium ion, phenol red composition), the child head of a family is to the bad acceptance of this method.And at present on the market the growth media of the cell of commercialized supply (GM), cell keep liquid (MM) and Eagle`s liquid (MEM), all contain microbiotic P.S solution (penicillin and Streptomycin sulphate), hypersensitive hand foot mouth disease infant is had a harm qualitatively.Hand foot mouth disease infant in addition, after tangible clinical symptom is arranged, major applications antibiotic therapy, the remaining microbiotic of sampling has certain influence to laboratory diagnosis.Adopting the throat swab sampling is to check the most reliable method of hand foot mouth disease virus.Being used for doing the cell cultivation in the laboratory separates, therefore, cell cultures " gold standard " that always detect for many years as hand foot mouth disease, cell cultures is all very high to equipment, technology, collection of specimens and the requirement of transporting etc., the mast cell who gathers from the oral cavity, syncyte and mucopurulent secretion thing should not be mixed with, too much impurity should not be contained.To gathering swab requirement is arranged also: should be cotton swab and should not be wooden swab, because wooden having wiped away may be suppressed viral growth.Sample must be placed in the special transhipment training base, and is freezing, is embedded in the cell cultures dish in 24 hours.On expense, equipment, technical elements all requires very high, therefore, expense and time are to influence the biggest obstacle that different medical unit hand foot mouth disease viral diagnosis method is used and promoted, and a lot of researchists have carried out many-sided exploration in the improvement of the method for sampling of hand foot mouth disease virus.In of the present invention and isotonic bacteria and virus mouth wash sampling liquid, have the effect of guiding clinical diagnosis and treatment, also significant for preventing to abuse antibacterials.

Background technology is present, and cell cultures is still diagnosis hand foot mouth disease virus infection reliable experiment method the most, traditionally cell cultures as " gold standard ".Determine that with the method for cell cultures the influence factor that the infection of hand foot mouth disease virus relates to 5 aspects all has a side effect qualitatively.The object of the present invention is to provide a kind of in and isotonic bacteria and virus mouth wash sampling liquid, compare with other class sample solution and to possess following characteristics: 1, prescription is simple, easily preparation, salable product can be prepared in the basic unit laboratory, the enterovirus that also can better preserve Picornaviridae (for example: 4,5,7,9,10,16 types of Coxsackie virus (Coxsackie virus) A group, 2,5 types of Coxsackie B virus group; Part Echo virus (ECHO-viruses) and enterovirns type 71).And utilizing aspect the PCR diagnostic techniques, having to be equal to HANKS liquid and preserve and transport result.In the hand foot mouth disease sampling standard that the Ministry of Health determines, regulation can utilize physiological saline to preserve liquid as sampling, now in physiology reason salt solution, add D-glucose again, foetal calf serum can provide part nutrition for virus, prolong preservation period, so the enterovirus that more can better preserve Picornaviridae reaches the purpose that improves the isolating positive rate of zoo virus.2, when gathering pharyngeal sample, available mode of gargling collects the sample sample of desired area more easily, samples without the throat swab mode, so sense of discomfort such as be difficult for causing vomiting.After child's mistake is swallowed, harmless because of the collutory composition is isoosmotic sugared salts solution, be easy to obtain child's cooperation and the child head of a family's acceptance.Foetal calf serum and glycine can neutralize the remaining microbiotic of sampling and conventional reagents relatively, and the recall rate height pollutes for a short time, and clinical application effect is satisfied with.

Summary of the invention main points of the present invention are to select suitable component, and rationally are mixed, through heat, dissolve, technology such as mixing, cooling, packing, high-temperature sterilization form a kind of in and isotonic bacteria and virus mouth wash sampling liquid.

The culture medium prescription that the present invention selects following (gram, the every 100ml distilled water of ml/) sodium chloride 8.0-0.85; D-glucose 0.8-1.2; Foetal calf serum 2-5ml; Glycine 0.2-0.4; Distilled water adds to 100ml; Transfer pH to 7.2-7.4.

Sodium chloride, D-glucose are the requisite materials of histiocytic body fluid equilibrium; Foetal calf serum, glycine are the requisite materials of histiocytic nutritive equilibrium; Want before foetal calf serum uses deactivation (56 ℃, 30min), to eliminate complement activity.Carry out sterility test simultaneously.High-quality serum should be for faint yellow, and transparent, no haemolysis does not have precipitation, and color is dark slightly after the deactivation.After adding foetal calf serum in the test, the pH of sample solution may also can change, so also can add adjust pH behind the calf serum earlier.After sample solution prepares, should extract a little earlier and put into culturing bottle,, whether pollution be arranged to detect nutrient solution in 37 ℃ of built-in 24～48hr of incubator.Each dosing amount is advisable about with two weeks, and a dosing is not too many, prevents nutritive ingredient (being mainly amino acid) loss, causes experiment loaded down with trivial details or pollute.Glycine is used to the remaining microbiotic of sampling that neutralizes simultaneously.

The preparation method of product of the present invention is:

1), deactivation (56 ℃ 30min), to eliminate complement activity, are carried out sterility test simultaneously will be carried out before the foetal calf serum use;

2), in proportion with sodium chloride, D-glucose, glycine heating for dissolving in distilled water, PH7.2-7.4 is transferred in cooling back, 115 ℃ of pressuresteam sterilizations, 20-30min is chilled to 50 ℃;

3), under aseptic condition, add aseptic foetal calf serum, mix, aseptic subpackaged, refrigerate standby.

The present invention has the following advantages: (1) prescription is simple, easily preparation, and salable product can be prepared in the basic unit laboratory; (2) clinical application effect is satisfied, has solved the throat swab mode and has sampled, the sampling difficulty of sense of discomfort such as easily cause vomiting; (3) isoosmotic sugared salts solution is harmless; (4) nutritious, can prevent microbiotic remaining in the sample, reach the purpose that improves the isolating positive rate of zoo virus.

The present invention is described in further detail below in conjunction with embodiment for embodiment:

Be configured according to following table institute column data (gram, the every 100ml distilled water of ml/) and described step thereof:

Prescription one:

Sodium chloride 8.0-0.85; D-glucose 0.8-1.2; Foetal calf serum 2.0-5.0ml; Glycine 0.2-0.4; Distilled water adds to 100ml; Transfer pH to 7.2-7.4.

Claims (2)

1. the present invention relates to a kind of in and isotonic bacteria and virus mouth wash sampling liquid, it is characterized in that having following prescription (gram, the every 100ml distilled water of ml/): sodium chloride 8.0-0.85; D-glucose 0.8-1.2; Foetal calf serum 2-5ml; Glycine 0.2-0.4; Distilled water adds to 100ml; Transfer pH to 7.2-7.4.

A claim 1 described in and the preparation method of isotonic bacteria and virus mouth wash sampling liquid, it is characterized in that having following step:

1), deactivation (56 ℃ 30min), to eliminate complement activity, are carried out sterility test simultaneously will be carried out before the foetal calf serum use;

2), in proportion with sodium chloride, D-glucose, glycine heating for dissolving in distilled water, PH7.2-7.4 is transferred in cooling back, 115 ℃ of pressuresteam sterilizations, 20-30min is chilled to 50 ℃;

3), under aseptic condition, add aseptic foetal calf serum, mix, aseptic subpackaged, refrigerate standby.