CN104561225A - A bacterial reverse-inoculation identification method in blood testing - Google Patents

Abstract

Through reverse-inoculation after suspected disease microorganisms are subjected to inoculated culture, different types of the microorganisms are distinguished by utilization of different growth characteristics and colony characteristics for the different types of the microorganisms after bacterial culture, and a suspected disease bacterium is selected and subjected to drug allergy testing so as to provide basis for a certain type of infection for clinicians. The method provided by the invention has advantages of simple experiments, low costs, easy operation and implementation, good repeatability, and the like, and overcomes limitation of traditional clinical examination to some extent.

Description

The method of bacterium reversal connection qualification in a kind of blood test

Technical field

The present invention relates to medical science disease microorganism authentication technique field, specifically provide simply a kind of and the method for bacterium reversal connection qualification in blood test accurately.

Background technology

The blood of human normal be do not have germy.After human body is subject to bacteriological infection, according to the difference by bacteriological infection degree, simply blood can be divided into microbemia, septicemia, pyemia etc. by the illness that bacteriological infection causes.When resistibility reduces, bacteriological infection human body generally first causes the inflammation of certain part, as furuncle, carbuncle, pneumonia, encephalitis etc.Medically the illness entering circulation of blood by local inflammation position because of bacterium is called microbemia.At this moment often have a small amount of bacterium and invade blood, if the systemic conditions of people is fine, this small amount of bacterium can be eliminated by the white corpuscle in the various antibody, phagocytic cell and the blood that produce in health.And if angioinvasive germ is a lot, toxicity is very strong, or due to human body overwork, malnutritive, severe trauma or have the reasons such as other diseases and resistibility is extremely low time, bacterium also can produce hazardous and noxious substances, with flow propagation to whole body by amount reproduction.At this moment patients is the serious toxicity symptom such as high heat, shiver with cold, headache, body temperature Chang Gaoda more than 40 DEG C.Meanwhile, bacterium can invade inflammation, the suppuration that each internal organs cause internal organs.If send out skin, various fash can be there is.The illness that after medically bacterium being entered blood, growth and breeding causes in blood calls septicemia.If after suppurative bacterium enters blood, not only breed in blood, cause some internal organs to form the bad or ypotension of abscess or even organ dysfunction, perfused tissue, this illness is then referred to as pyemia.Sepsis case fatality rate is higher, and septic shock and many organ pipes functional defect can occur the sepsis patient of nearly 9%, and death over half in intensive care unit is caused by septic shock and multiple organ dysfunction.Research shows, occur internal organs organ failure, shock, multiple infection, serious potential disease patient's prognosis poor.A common ground of these three kinds of illnesss of above microbemia, septicemia, pyemia has bacterium in blood samples of patients.If cultivated by blood samples of patients, namely pathogenic bacterium can be detected.Therefore, the Clinics and Practices of result to disease of blood microbial culture and qualification is significant.

In general, the sample of blood microbial culture is gathered by reviewer, and the sample process of collection is hatched, culture transferring, tentatively judges, and then sends report according to the Identification of Biological Characteristics of various pathogenic bacteria.But in actual clinical inspection, the blood sample of collection directly hatches cultivation, usually there will be the situation that some specific pathogenic bacterium fails to detect.Especially in recent years, blood infection occurs in rising trend, the good hospital of many appointed conditions substantially all adopts the equipment of automatic blood microbial culture to carry out blood microbial culture, the positive rate that there is hemoculture is at large not high enough, culture cycle is long, easily occur that pollution leads to errors the problems such as result, this all has very tremendous influence to selecting medication, timely and effectively infection control and last patient effect in clinical treatment.

Therefore provide simply a kind of and in blood test accurately bacterium reversal connection qualification method there is extremely important meaning.

Summary of the invention

Produce for existing the technical problem existed, the object of this invention is to provide a kind of sorting picking method of pickle disease bacterium simple to operate, safe and reliable, with low cost.

The inventive method, according to the conspicuous rule of section, adopts traditional microbiological authentication method and hematology testing method to combine.Concrete technical scheme is the method for bacterium reversal connection qualification in a kind of blood test, it is characterized in that comprising step:

1) sampling is cultivated: sample from blood sample with aseptic technique and carry out pathogenic bacteria and hatch, and is placed in shaking table cultivation 10 ~ 24h;

2) reversal connection is cultivated: get liquid level upper liquid as sample, smear reversal connection on blood serum medium, be inverted in constant temperature culture 2 ~ 3d in 36 DEG C of constant incubators, observes colony growth situation;

3) preliminary susceptibility: picking disease bacterium, preliminary antibiotics susceptibility test;

4) identify: identify according to assay, optionally carry out secondary antibiotics susceptibility test.

The present invention cultivates by carry out after inoculation culture reversal connection again to doubtful disease microorganism, the growth characteristics that after utilizing microbial culture, different types of microorganism is different and colony characteristics make it be distinguished and choose the doubtful bacterium of disease to carry out antibiotics susceptibility test, for clinician provides foundation for a certain specific infection.The inventive method has experiment enforcement simple, with low cost, easy to operate, the advantage such as reproducible, can overcome the limitation of traditional clinical inspection to a certain extent.

Embodiment

Below with reference to specific embodiment, the present invention is described in further details.

Embodiment

Test raw material and reagent

1) blood sample of patient;

2) serum-based basal culture medium: serum homogeneous heating to 56 DEG C is kept 30min, and be stored in 4 DEG C, mix afterwards with minimum medium, minimum medium accounts for 80% ~ 90%, and serum accounts for 10% ~ 20%.Dual anti-stock solution (penicillin+Streptomycin sulphate) is added by 1% volume fraction;

3) antibiotics susceptibility test: piperacillin sodium-tazobactam, gentamicin, vancomycin, clindamycin, Cefuroxime sodium, cefotaxime sodium.

Testing sequence:

1) sampling is cultivated: sample from blood sample with aseptic technique and carry out pathogenic bacteria and hatch, and is placed in shaking table cultivation 10 ~ 24h;

2) reversal connection is cultivated: get liquid level upper liquid as sample, smear reversal connection on blood serum medium, be inverted in constant temperature culture 2d in 36 DEG C of constant incubators, observes colony growth situation;

3) preliminary susceptibility: picking disease bacterium, preliminary antibiotics susceptibility test;

4) identify: identify according to assay, optionally carry out secondary antibiotics susceptibility test.

Claims (1)

1. a method for bacterium reversal connection qualification in blood test, is characterized in that comprising step:

1) sampling is cultivated: sample from blood sample with aseptic technique and carry out pathogenic bacteria and hatch, and is placed in shaking table cultivation 10 ~ 24h;

2) reversal connection is cultivated: get liquid level upper liquid as sample, smear reversal connection on blood serum medium, be inverted in constant temperature culture 2 ~ 3d in 36 DEG C of constant incubators, observes colony growth situation;

3) preliminary susceptibility: picking disease bacterium, preliminary antibiotics susceptibility test;

4) identify: identify according to assay, optionally carry out secondary antibiotics susceptibility test.