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Disinfection technical specifications2002(3)

PubDate:2015/9/22 10:44:48   Category:News [Print][Close]

 2.1.2 disinfectant simulation and field disinfection test on site

2.1.2.1 disinfectant simulation of food (drink) with disinfection effect evaluation test on site

2.1.2.1.1 purpose

Identification of disinfectant of killing bacteria on the food (drink), to verify that the disinfectant of food (drink) with disinfection practical dose.

2.1.2.1.2 test equipment

E. coli (1) (8099). Is still needed to kill other specific bacteria, but increase with the specific bacteria tested. According to provisions of 2.1.1.2 bacteria suspension preparation methods and requirements.

(2)) phosphate buffer solution (PBS, 0.03 mol/L, pH7.2)

(3) the diluent: see appendix A.

(4) the neutralization agent (according to 2.1.1.5 methods identify eligible)

(5) tryptone soy AGAR medium

(6) sterile cotton swabs

(7) specifications plate (preparation with soft materials can be bent slightly, the central one size is 5.0 cm x 5.0 cm space as sample parts).

(8) test with food (drink) with sample bowls (plate) or bamboo (wood) chopsticks front-end (circumference of 2.0 cm, 12.5 cm in length.

2.1.2.1.3 operating procedures

(1) according to the use of the product specification, selected the test employs concentration and action time, kill e. coli tests are carried out respectively.

Sterile specification plate (2) using in test serve (disc) mark dye intermediate zone (5.0 cm x 5.0 cm). Drops separately with the aseptic suction drain bacteria suspension, add in dye bacterium area, were 0.1 ml, with sterile L bar in the area with dry 37 ℃ constant temperature box.

To take chopsticks front-end 12.5 cm length, dip dyeing after booking volume bacteria liquid, and 37 ℃ or room temperature dry.

(3) experimental group: 30 dyed sample bowl or 3 0 chopsticks, in turn, timing is put into the container containing disinfectant solution, so that it is completely submerged. Role to a certain time, the rejected disinfectant, the bowls (plate) within the dyed area adding 5 ml neutralizer, new-developped blow wash with a L rod bacteria, after 10 min, 1.0 ml samples were taken into 2 pieces of plate. Put chopsticks in containing 20 ml ~ 25 ml of neutralizer test tubes, make its total immersion in the neutralizer, electric vortex mixer shock 60 s or vibration on 200, after 10 min, respectively take 1.0 ml sample liquid pour into 2 pieces of plate. Put 37 ℃ constant temperature box the vaccination good AGAR culture 48 h, number counting colonies, as a group.

(4) positive control group: the three dye bacterium bowl or 3 samples of chopsticks don't soak disinfectants, directly to the bowls (plate) within the dyed area adding 5 ml neutralizing agent, or directly put chopsticks in containing 20 ml ~ 25 ml neutralizer test tubes. After being group disinfection processed, sampling and testing in accordance with the experimental group. Positive control bacteria amount should be 1.25 ~ 1.25 x 108 x 107 cfu/sample cfu/sample (the equivalent of 5 x 105 cfu/cm2 ~ 5 x 106 cfu/cm2).

(5) the negative control group: treated group and positive control group after the test, the unused with batch neutralizer, PBS inoculated medium and kinds of bacteria culture medium and so on of the above two sets of samples for culture at the same time, as a negative control.

(6) press type calculation tableware per piece on the growth of colony count.

Tableware per sample number growth colonies (cfu/sample) = tablet on the average number of colonies by testing the sample dilution multiple

(7) to kill the value.

2.1.2.1.4 evaluation rules

For each type of food (drink) with 30 samples of e. coli to kill for numerical acuity 3.00 concentration and action time of the disinfectant used for disinfection qualified dose.

2.1.2.1.5 matters needing attention

(1) test process must take strict aseptic technique. Direct or indirect contact with the sample of the equipment must be used after sterilization. Simulated field test used in the food (drink) with before dyeing bacteria also must undergo processing.

(2) the positive and negative control must be set for each test, and never can be omitted.

2.1.2.2 disinfectant disinfection simulated field test of medical devices

2.1.2.2.1 purpose

Identification of disinfectant of artificial pollution in bacterial spore killing effect on medical apparatus and instruments, to verify that the processing of medical devices and practical dose.

2.1.2.2.2 test equipment

(1) black variant (ATCC 9372) spore bacillus subtilis (hereinafter referred to as spores, the preparation of the suspension by 2.1.1.2.3 (2) the provisions)

(2) phosphate buffer solution (PBS, 0.03 mol/L, pH7.2)

(3) the neutralizer solution (via methods as stipulated in the 2.1.1.5 appraisal eligible)

(4) tryptone soy broth culture medium containing neutralizing agent

(5) to simulate the field experiment with samples (for medical hemostatic forceps truncation, take its from shaft to gear tooth part, reference 2.1.1.2.4 defatted processing method (2) regulations. After pressure steam sterilization, dried standby).

(6) plastic tube (1.8 cm by 18 cm).

2.1.2.2.3 operating procedures

(1) to carrier immersion quantitative germicidal test to determine the effect of disinfection. According to the selected product specification dose, test employs concentration and action time.

(2) to dye the fungus, the hemostatic forceps with sterile tweezers samples on the tooth face, and fixed on the sterile supports. With 0.1 ml sterile sterile liquid move straw or quantitative, will 0.02 ml spore suspension droplet dye in the tooth, with sterile L with platinum wire, 37 ℃ constant temperature drying in the standby.

(3) take asepsis AGAR, dosage of 10 ml per carrier to join a detergent, 20 ℃ insulation plus or minus 2 ℃ water bath in 5 min.

(4) will be 30 dye bacterium samples immersed in a disinfectant in disinfection treatment.

(5) the role to the specified time, remove samples, respectively into 10 ml of the neutralizer solution plastic in vitro. Palm vibration on 200 times, respectively 1.0 ml sample liquid poured vaccination two asepsis AGAR, put 37 ℃ constant temperature box to develop 72 h, number counting colonies, as a group.

(6) replace the disinfectant with sterile distilled water, the three dye sample processing in the same conditions, then living bacterium culture method with experimental sample count, as the positive control group, the bacteria amount should be in 5 ~ 5 x 106 x 105 cfu/sample cfu/samples.

(7) after the test, will use the same batch of neutralizing agent, sample liquid culture medium inoculated with diluent, training; The other will not used the same batch of medium is also in constant temperature box culture, as a negative control.

2.1.2.2.4 evaluation rules

Within the prescribed period of time, positive control group were bacterial growth, the number of living bacterium culture count reaches, negative control are sterile growth, in the experimental group 30 samples on artificial pollution spore killing rate of the logarithmic average of 3.00 or higher, can be jailed for disinfection.

2.1.2.2.5 matters needing attention

In connection with this test the matters of attention in the other test is also suitable for this test.

2.1.2.3 disinfectant simulated field sterilization test of medical devices

2.1.2.3.1 purpose

Is used to validate disinfectant of artificial contaminated with spores of medical device sterilization effect.

2.1.2.3.2 test equipment

(1) black variant (ATCC 9372) spore bacillus subtilis (hereinafter referred to as spores, the preparation of the suspension by 2.1.1.2.3 (2) the provisions)

(2) phosphate buffer solution (PBS, 0.03 mol/L, pH7.2)

(3) the neutralizer solution (via methods as stipulated in the 2.1.1.5 appraisal eligible)

(4) tryptone soy broth culture medium containing neutralizing agent

(5) to simulate the field experiment with samples (for medical hemostatic forceps truncation, take its from shaft to gear tooth part, reference 2.1.1.2.4 defatted processing method (2) regulations. After pressure steam sterilization, dried standby).

(6) plastic tube (1.8 cm by 18 cm)

2.1.2.3.3 operating procedures

(1) according to the product specification 0.5 times the minimum use of concentration and the shortest duration.

(2) 25 dye the bacteria in the sample preparation by 2.1.2.2.3 (2) the provisions.

(3) sterile AGAR, according to the dosage of 10 ml each carrier to join a detergent, 20 ℃ + 2 ℃ water bath for insulation in 5 min.

(4) to 20 dye bacterium samples immersed in disinfectant sterilization processing, effect to the provisions of time, remove samples, respectively in 10 ml of neutralizer broth (20) in a test tube, placed in 37 ℃ incubator qualitative 7 d, observe the final result, as a group. With bacteria growth, a mild broth medium turbidity, European bacteria biofilm, vibration wave gently visible floc, sterile broth of transparent growth.

(5) in a standard hard water instead of disinfectant, the two dyeing carrier to handle under the same conditions, and then with experimental group carrier method of inoculation, cultivation, observation, as the positive control group.

Taken 3 (6) the other dye bacteria samples, respectively in a 10 ml neutralizer solution plastic in vitro. Oscillation 1 min, in the palm or vibration on 200 times, sampling liquid with live bacteria count. 72 h counting colonies. As the number of control group, the bacteria amount shall be the 5 ~ 5 x 106 x 105 cfu/sample cfu/samples.

(7) after the test, will use the same batch of neutralizing agent, liquid sampling and diluent to vaccinate medium for culture, the other will not use the same batch of medium is also in constant temperature box culture, as a negative control.

(8) test repeat 3 times.

2.1.2.3.4 evaluation rules

All 60 samples are sterile growth, all test group and positive control group have the bacteria growth, the number controls the number of living bacterium culture count reaches, negative control group are sterile growth, can be judged to be qualified sterilization.

2.1.2.3.5 matters needing attention

(1) sterilization effect observation, strict aseptic operation, otherwise qualified sterilization can be mistaken for failure.

(2) in this experiment on the matters of attention in the other test is also suitable for this test.

2.1.2.4 continuous use stability test

2.1.2.4.1 purpose

Verify the disinfectant soaking in repeatedly take put the use of medical equipment under the condition of the period of validity.

2.1.2.4.2 test equipment

(1) black variant (ATCC 9372) spore bacillus subtilis (hereinafter referred to as spores, the preparation of the suspension by 2.1.1.2.3 (2) the provisions)

(2) phosphate buffer solution (PBS, 0.03 mol/L, pH7.2)

(3) the neutralizer solution (via methods as stipulated in the 2.1.1.5 appraisal eligible)

(4) tryptone soy broth culture medium containing neutralizer: see appendix A.

(5) test samples [reference 2.1.1.2.4 (2)].

(6) carrying medical devices (choose surgical scissors, hemostatic forceps, such as small instruments, or choose other instrument according to need.

(7) sterile with cover enamel disc.

2.1.2.4.3 operating procedures

(1) black with bacillus subtilis variant spore bacteria for test.

(2) test, make up a double 2000 ml disinfectant, sterile with cover are dressed in two enamel plate. Each into a clean test with medical apparatus and instruments, make up to full load requirements. Each test, at the same time, respectively, to test the two enamel pan disinfectant.

(3) after enamel disc into the instrument, daily remove equipment, washing with water, drain and then play back into the disinfectant. Continuous daily remove equipment, washing, drain well, put a, until the end of the test.

(4) into the instrument to the directions of continuous use for the longest time, and cancel the venom samples, disinfection of medical equipment as disinfectant for medical instrument disinfection simulated field test (2.1.2.2), sterilization of medical equipment as disinfectant for medical device simulation field sterilization experiment (2.1.2.3), the method of determination of the disinfectant effect of killing spores.

2.1.2.4.4 evaluation rules

Repeat 3 times to 3 times test results meet the requirements of qualified, can be jailed for continuous use stability test.

2.1.2.4.5 matters needing attention

(1) the disinfectant enamel disc into the, shall be affixed to each time after the operation.

(2) the sterilization effect of observation, strict aseptic operation, otherwise qualified sterilization can be mistaken for failure.

(3) the test about the matters of attention in the other test is also suitable for this test.

2.1.2.5 disinfectant disinfection simulated field test

2.1.2.5.1 purpose

Used for testing the disinfectant of artificial pollution surface bacteria killing effect on hand, and for sure this disinfectant disinfection practical dose reference.

2.1.2.5.2 test equipment

E. coli (1) 8099 or NCTC 10538, is still needed to kill other bacteria specific purpose, can increase with the specific bacteria tested. According to provisions of 2.1.1.2 bacteria suspension preparation methods and requirements.

(2) tryptone soy AGAR (TSA), see appendix A.

(3) tryptone soy broth culture medium (TSB) : see appendix A.

(4) the neutralizer (TSB as the solvent, the accreditation).

(5), 200 g/L liquid soap.

(6) reference sample: propyl alcohol 60% 60% (V/V) with isopropyl alcohol (V/V).

(7) standard hard water, see appendix A.

(8) phosphate buffer solution (PBS, 0.03 mol/L, pH7.2).

(9) sterile gauze cloth.

(10) pipe, tube, plate number.

(11) oscillation mixer.

2.1.2.5.3 test steps

(1) the preparation of bacterial suspension in TSB culture take two of 18 ~ 24 h h e.coli cultures are vaccinated in 1 LTSB triangle flask, in 36 ℃ + / - 1 ℃ cultivation in 18 ~ 24 h h, dilute it with the TSB to 2 x 108 cfu/ml ~ 2 x 109 cfu/ml bacteria suspension.

(2) to 12 ~ 15 volunteers were randomly divided into two groups, the first group to use the reference samples, the second group used test samples

(3) using liquid hand soap for 1 min to remove surface contamination of the natural bacteria, then hand wipe with a sterile gauze.

(4) will be 2 x 108 cfu/ml - 2 x 109 cfu/ml of e. coli bacteria suspension into a sterile containers,

(5) the hands to the tip of the finger parts into bacteria suspension, fingers apart, keep 5 s, will leave bacteria suspension, hand in the air dry for 3 min.

(6) dry immediately after your thumb and other fingers in a petri dish containing 10 ml TSB rub for 1 min, take appropriate vaccination AGAR dilution degrees. Number counting colonies, as bacteria number before test.

(7) hands, not repeat pollution disinfection treatment immediately.

(8) test sample groups: according to the instructions, the dosage of action time and the use of the frequency in a standard way to wash your hands (as shown in figure 2-1) rub the 30 s to 60 s longest (health hand disinfection) or 5 min (surgical hand disinfection). To the flow of tap water rinse 5 s, shake off the hand of residual water. Immediately your thumb and other fingers in a petri dish containing 10 ml neutralizer rub for 1 min, make appropriate diluted, respectively 1.0 ml sample liquid to pour into two asepsis AGAR method immunization, put 37 ℃ constant temperature box cultivate 48 h, number counting colonies.

Reference sample group (9) : for health hand disinfection, take 60% isopropyl alcohol 3 ml to start in the heart, according to the standard way to wash your hands hard rub the 30 s. In order to ensure that all parts of the hand contact the isopropyl alcohol. Repeated use of isopropyl alcohol rub as above make the total rub the time for 60 s. For surgical hand disinfection, using 60% normal propyl alcohol 3 ml, according to the standard way to wash your hands scrub brush, when nearly dry, add 3 ml is propyl alcohol rub is brushed. In order to keep 3 min. With flowing water flushing of the 5 s, shake off the remaining water. The rest of the steps the same as the test sample set.

 

 

 

 

 

 

 

1. The palm to palm rub the 2. Fingers interlaced palm opponent back rub is brushed 3. Fingers interlaced palm to palm rub is brushed

 

 

 

 

 

 

Rub them clasped each other 4. Refer to back 5. Thumb rotation rub the 6. In the long fingers in the palm of friction

Figure 2-1 the standard way to wash your hands

(10) in the test on the same day, the first group of exchange with the second sample, repeat the test.

(11) to calculate the reference samples and test samples group on the number of bacteria decrease the value.

2.1.2.5.4 evaluation rules

(1) test samples of kill kill of numerical value is greater than or equal to the reference samples for value as a qualified.

(2) if the test samples to kill to kill the numerical value is less than the reference sample, should carry on the statistical treatment, to determine whether the difference is remarkable.

(3) if the test samples of killing of killing no significant value is less than the reference sample for value, can be jailed for disinfection. If test samples of kill to kill to numerical value significantly less than the reference sample, show that the test sample is not conform to the requirements of this specification.

2.1.2.5.5 matters needing attention

(1) the test must be in the same subjects, on the same day, the same environment, under the same conditions, make the results of the test sample and reference sample are comparable.

(2) all the participants should be at least 18 years old, healthy body. Hand skin should be without damage, without skin, nails short and clean.

(3) test bacteria suspension should be within 3 h after 1 hand in use. In one experiment, using the test sample or reference sample, all subjects of hand dye bacterium should use the same bacteria suspension.

(4) of the subjects, at the end of each test, should be timely and thorough disinfection treatment on hands.

2.1.2.6 disinfectant disinfection field test

2.1.2.6.1 purpose

Determination of disinfectant opponent surface disinfection of natural bacteria needed to use dose, to verify that the disinfectant disinfection practical dose.

2.1.2.6.2 test equipment

(1) the neutralizer (neutralizer identified qualified)

(2) sterile cotton swabs

(3) the diluent 0.1% twain 80 0.03 mol/L pH7.2 phosphate buffer

(4) phosphate buffer solution (PBS, 0.03 mol/L, pH7.2).

(5) standard hard water, see appendix A.

(6) sterile gauze cloth

(7) tryptone soy broth culture medium (TSB) : see appendix A.

(8) tryptone soy AGAR (TSA), see appendix A.

(9) oscillation mixer

(10) pipe, tube, plate number.

2.1.2.6.3 test steps

(1) in the use of the scene, randomly selected subjects. Test not less than 30 people.

(2) the disinfection before, after the subjects both hands fully rub each other, let the participants left hand together, with sterile cotton swabs soaked in 10 ml diluent tube, on the wall after drying, the finger flexor fingertips to root, inunction 2 times back and forth, each inunction again, cotton swabs of turning once more. After sampling, with sterile operation mode will be cut with the cotton swabs of sampling neutralizer in vitro, as the positive control sample.

(3) according to the instruction manual method was carried out on the right hand disinfection disinfectant, the opponent's health disinfection generally set the duration of 1 min, for surgeons to wash their hands after hands in general the duration to 3 min. After disinfection by neutralizing agent instead of diluent, the same way as the positive control group of subjects on the right hand the remaining natural bacteria sampling time, as experimental samples.

(4), respectively, will not use the same batch of neutralizing agent, diluent, 1.0 ml, cotton swabs of 1 ~ 2 samples as the negative control group.

(5) respectively in trial group, positive control group and the negative control group 1 ml sample, with inoculation of AGAR AGAR pouring method, each sample vaccination two plate, put 37 ℃ constant temperature box in the cultivation of 48 h, observe the final result.

(6) to kill the value

2.1.2.6.4 evaluation rules

Positive control group should be more bacterial growth and the negative control group should be sterile growth, to 30 m on average the natural bacteria killing of numerical acuity 1.00 can be jailed for disinfection.

2.1.2.6.5 matters needing attention

(1) the test should be volunteers, repeat visitors is more, a person can be subjects for many times, but not in a batch test or repeated on the same day, otherwise can affect the accuracy of the results.

(2) subjects were tested, shall not touch any surface, lest make test part of the hand with mixed bacteria.

(3) cotton swabs daub, sampling, difficult to standardize, therefore should try to make the size of the cotton swabs, force uniform, absorb the quantity of sample liquid, and wash the bacteria was the weight of the hammer made consistent.

(4) to wipe disinfection, tu dose to appropriate, apply evenly.

(5) the sample must be timely detection, stored at room temperature shall not exceed 2 h. Otherwise should put 4 ℃ refrigerator, but also to no more than 4 h.

2.1.2.7 disinfectant for skin disinfection simulated field test

2.1.2.7.1 purpose

Determination of disinfectant on the skin surface bacteria killing effect of artificial pollution, to verify the disinfectant for skin disinfection practical dose.

2.1.2.7.2 test equipment

(1) test strains: staphylococcus aureus ATCC 27217, is still needed to kill other specific bacteria, but increase with the specific bacteria tested.

(2) tryptone soy AGAR (TSA), see appendix A.

(3) phosphate buffer solution (PBS, 0.03 mol/L, pH7.2).

(4) the neutralizer (neutralized agent accreditation).

(5), 200 g/L liquid soap.

(6) metal tube (2.2 cm in diameter, height of 3 cm).

(7) nylon scraping bacteria stick.

(8) sterile gauze cloth.

(9) pipe, tube, plate number.

(10) oscillation mixer.

(11) one-time inoculation loops (4 mm in diameter).

(12) antibiotic ointment.

(13) skin disinfectant.

Constant temperature box (14).

2.1.2.7.3 test steps

(1) preparation of bacteria suspension by 2.1.1.2.3 regulation methods and requirements.

(2) use liquid soap cleaning subjects forearm medial 1 min, use soap water rinse residues, and then use dry sterile gauze, the natural bacteria to remove the forearm medial surface contamination.

(3) with one end filling ink is 3.0 cm in diameter glass tube buckles on the subjects of each forearm midway (don't) at the wrist and elbow creases, divided into an experimental zone.

Washer (4) use take 10 mu l the bacteria suspension, vaccination in the forearm test area (the recovery of colony count to 2 x 106 cfu/test area ~ 1 x 107 cfu/test area), the use of disposable inoculation loops, the bacteria suspension painted a circle, and the experimental zone edge should be 4 mm to 5 mm distance, natural dry in the air.

(5) according to the instruction for use disinfectant method was carried out on the right forearm medial disinfection, generally set the duration of 1 min ~ 3 min.

(6) after disinfection by neutralizing agent of forearm vaccination to sampling area. When sampling, the metal tube placed in the experimental zone in the middle part, the area covered with bacteria, carries don't come into contact with the edge of the ink. Remove the 1.0 ml neutralizer to metal tube, nylon scraping bacteria stick a wash metal cylinder covered area of 60 s skin, remove the absorption liquid in the cylinder to in vitro, plus 1.0 ml neutralizer to a second blow in the area of skin wash 30 s, will be the second scrubbing liquid, injection test tube containing the first blow wash liquid, as experimental samples.

(7) in distilled water instead of disinfectants used to do the same with the middle of the left forearm as control sample.

(8) respectively for each test will not used the same batch of neutralizing agent, diluent, 1.0 ml sample as the negative control group.

(9), respectively, to take appropriate sample dilution degree of test group and control group and the negative control group samples each 1.0 ml, with inoculation of AGAR AGAR pouring method, each sample vaccination two plate, put 37 ℃ constant temperature box in the cultivation of 48 h, observe the final result.

(10) kill for numerical calculation.

(11) test shall not be less than 15 m.

2.1.2.7.4. Evaluation rules

Control group recycling colony count of 2 x 106 cfu/test area ~ 1 x 107 cfu/test area, the negative control group should be sterile growth, and to each people artificial skin surface contamination of staphylococcus aureus to kill the numerical are 3.00 or higher can be jailed for disinfection qualified.

2.1.2.7.5 matters needing attention

(1) the subjects recruitment standards

1) ranging in age from 18 to 65 - year - old male or female;

2) subjects should be in good health;

3) the forearm skin should be in good condition and no skin and other skin problems.

(2) the rule out the standard of the subjects, if subjects had one of the following circumstances, cannot be employed to participate in the test

1) pregnant women;

2) diagnosed with diabetes, hepatitis, HIV/AIDS (HIV), a transplant.

(3) after the test sample, with 70% of alcohol on subjects forearm disinfected first, and then use the skin disinfectant for two forearm disinfection treatment, treatment after wash, dry, then apply a small amount of antibiotic ointment in the experimental area, in case of skin infections.

(4) after completion of the test within 48 h, 72 h, the subjects such as found a pimple on the forearm, blisters, of raised, red itchy blister shall timely notify the inspection unit.

2.1.2.8 disinfectant for skin disinfection field test

2.1.2.8.1 purpose

Determination of disinfectant in disinfection of natural bacteria on the skin surface the dosage required to verify the disinfectant disinfection practical dose.

2.1.2.8.2 test equipment

(1) the neutralizer (neutralizer accreditation).

(2) sterile cotton swabs.

(3) the diluent 0.1% twain 80 0.03 mol/L pH7.2 phosphate buffer.

(4) phosphate buffer solution (PBS, 0.03 mol/L, pH7.2).

(5) standard hard water, see appendix A.

(6) sterile gauze cloth.

(7) specifications plate (preparation with kraft paper, the central leave a 3.0 cm by 10.0 cm space as a sampling site) 121 ℃ for 15 min sterilization. Set aside.

(8) tryptone soy AGAR (TSA).

(9) oscillation mixer.

(10) pipe, tube, plate number.

2.1.2.8.3 test steps

(1) in the use of the scene, randomly selected subjects. Test not less than 30 people.

(2) before disinfection, subjects, left and right forearm medial middle after fully to rub each other, put on the left forearm midway through the inside of the subjects the specification plate surface, using sterile cotton swabs soaked in 10 ml diluent tube, on the wall after drying, in the area of specification plate frame, lateral inunction 10 times back and forth, longitudinal inunction, 3 times back and forth every inunction again, cotton swabs of turning once more. To aseptic operation after sampling, cotton swabs of sampling side cut into the sample liquid in vitro, shaking 200 times.

(3) according to the disinfectant use manual method was carried out on the right forearm medial disinfection, generally set the duration of 1 min ~ 3 min. After disinfection by neutralizing agent instead of diluent, the same way as the positive control group of the subjects right forearm medial surface sampling time, the remaining natural bacteria as experimental samples.

(4), respectively, will not use the same batch of neutralizing agent, diluent and cotton swabs samples as the negative control group.

(5) respectively take appropriate dilution degree of test group, positive control group and the negative control group samples each 1.0 ml, with inoculation of AGAR AGAR pouring method, each sample vaccination two plate, put 37 ℃ constant temperature box in the cultivation of 48 h, observe the final result.

(6) to kill the value

2.1.2.8.4 evaluation rules

Positive control group should have more bacterial growth, negative on group should be sterile growth, to over 30 batches of the natural bacteria on the skin surface to kill for numerical acuity 1.00, can be jailed for disinfection.

2.1.2.8.5 matters needing attention

(1) the test should be volunteers, repeat visitors is more, a person can be subjects for many times, but not in a batch test or repeated on the same day, otherwise can affect the accuracy of the results.

(2) subjects were tested, shall not touch any surface, lest make test parts with mixed bacteria.

(3) cotton swabs daub, sampling, difficult to standardize, therefore should try to make the size of the cotton swabs, force uniform, absorb the quantity of sample liquid, and wash the bacteria was the weight of the hammer made consistent.

(4) to wipe disinfection, tu dose to appropriate, apply evenly.

(5) the sample must be timely detection, stored at room temperature shall not exceed 2 h. Otherwise should put 4 ℃ refrigerator, but also to no more than 4 h.

2.1.2.9 disinfectant to other surface disinfection simulation evaluation test on site

2.1.2.9.1 purpose

Used to identify disinfectant for artificial pollution from general object [in addition to this specification has special provisions, such as food (drink), medical equipment and other objects] the killing effect of the bacteria, to verify the disinfectant on the surface disinfection of practical dose.

2.1.2.9.2 test equipment

(1) (8099) and staphylococcus aureus, e. coli (ATCC 6538). If you need to kill their specific microorganisms can increase with the specific microbial test. According to provisions of 2.1.1.2 bacteria suspension preparation, methods and requirements, microbial testing should be in 1.25 ~ 1.25 x 108 x 107 cfu/sample cfu/sample (the equivalent of 5 x 105 cfu/cm2 ~ 5 x 106 cfu/cm2).

(2) phosphate buffer solution (PBS, 0.03 mol/L, pH7.2)

(3) the diluent (80 PBS solution containing 0.1% twain)

(4) the neutralization agent (according to 2.1.1.5 methods identify eligible)

(5) tryptone soy AGAR medium

(6) sterile cotton swabs

(7) specifications plate (stainless steel material preparation, the central leave a 5.0 cm by 5.0 cm space as a sample)

2.1.2.9.3 test steps

(1) with artificial dye fungus material such as desktop, ground, wall for disinfection. Under the condition of no special requirements, can use wooden desktop, disinfection effect is observed.

(2) each test, all kinds of surface testing 30 samples.

(3) to dye the fungus, the selected item is flat area, within the specification plate central space using sterile cotton swabs dipped in the bacteria suspension evenly temperature is below 60 block (25 cm2). After natural drying experiment. 30 blocks as a positive control area, 30 blocks for test area.

(4) positive control group: the sterile cotton mop in a test tube containing 5 ml diluents wet and in 30 control block daub sampling, each block bearing round-trip eight times. After sampling, with sterile operation mode will cotton swabs sample cut into the original diluent in vitro, vibration to play 200 times, 10 min. Do appropriate dilution with diluent.

(5) group: according to the instructions described in the use of doses to disinfectant spray or paint to the surface disinfection. After disinfection, sterile cotton swabs to contain 5 ml wet neutralizer test tubes, respectively to daub sampling of disinfection of 30 blocks, each block bearing round-trip eight times. After sampling, with sterile operation mode will cotton swabs sample cut into the original neutralizer in vitro, vibration to play 200 times, 10 min. Neutralizing agent are used when necessary for appropriate dilution.

(6) after the test, will use the same batch of 1.0 ml inoculated medium, neutralizing agent and diluent as negative control samples.

(7) to positive control, negative control group and disinfect group samples, each draw 1.0 ml, with inoculation of AGAR AGAR pouring method, each sample vaccination two plate, put 37 ℃ constant temperature box in the cultivation of 48 h, observe the final result.

(8) kill for numerical calculation.

2.1.2.9.4 evaluation rules

Repeat 3 times, number of positive control group conform to the requirements, negative on groups of sterile growth, all kill of numerical samples are 3.00 or higher, can be jailed for disinfection.

2.1.2.9.5 matters needing attention

(1) test operation must take strict aseptic technique.

(2) must be set for each test positive and negative control, never can be omitted.

(3) before and after disinfection, sampling (positive control group and disinfection trial group), shall not be in the same area.

(4) cotton swabs daub sampling are difficult to standardize, therefore should try to make the size of the cotton swabs, force uniform, absorb the quantity of sample liquid, washed bacteria was the weight of the hammer and so on successively.

(5) the sample must be timely detection. Room temperature storage must not exceed 2 h, otherwise should be 4 ℃ refrigerator, but also must not exceed 4 h.

2.1.2.10 disinfectant for other surface disinfection field evaluation test

2.1.2.10.1 purpose

Used to identify disinfectant for general object [in addition to this specification has special provisions, such as food (drink), medical equipment and other objects] the killing effect of the natural bacteria, to verify that the disinfection disinfectant on the surface of the practical dose.

2.1.2.10.2 test equipment

(1) phosphate buffer solution (PBS, 0.03 mol/L, pH7.2)

(2) the neutralizer

(3) the diluent (80 PBS solution containing 0.1% twain)

(4) sterile cotton swabs

(5) specifications plate (stainless steel material preparation, the central leave a 5.0 cm by 5.0 cm space as a sample)

(6) tryptone soy AGAR medium

2.1.2.10.3 operating procedures

In the use of the scene, according to the instructions of the dosage, duration, frequency of use and disinfection methods disinfection surface, a test sample should be 30 or more.

(1) random take surfaces (desktop, mesa, door, etc.), with specification calibration 2 boards area is 25 cm2 blocks, each one for sampling before disinfection, a sampling after disinfection.

(2) before disinfection, sterile cotton swabs will be in a test tube containing 5 ml diluents wet, daub sampling in one area, namely between the eight times. To aseptic operation after sampling, cotton swabs sample cut into the original diluent in vitro, vibration shock 20 s or to play 80 times, make appropriate diluted, as the positive control sample.

(3) according to prescribed dose, the disinfectant spray or paint to the surface disinfection. After disinfection, sterile cotton swabs to contain 5 ml wet neutralizer test tubes, the disinfection block daub sampling, namely between the eight times. To aseptic operation after sampling, cotton swabs of sampling cut into the original sampling fluid in vitro, electric vortex mixer shock 20 s or vibration to play 80 times, as disinfection group samples.

(4) after the test, will use the same batch of 1.0 ml inoculated medium, neutralizing agent and diluent as negative control samples.

(5) the positive control group, negative control group and disinfect group samples, each draw 1.0 ml, with inoculation of AGAR AGAR pouring method, each sample vaccination two plate, put 37 ℃ constant temperature box in the cultivation of 48 h, observe the final result.

(6) to kill the number value.

2.1.2.10.4 evaluation rules

Repeat 3 times, positive control group should have more bacterial growth, negative growth of group should be sterile, disinfection sample average kill out of numerical 1 or more, can be jailed for disinfection.

2.1.2.10.5 precautions:

(1) in the field test, the natural bacteria species complex, on the tablet in large area of mold growth and lead to can't counting colonies. At this time, in two parallel plate, such as a tablet to count colony count, as the plate colony counting results. As both tablet have mold growth and large area should be tested again.

(2) the test operation must take strict aseptic technique.

(3) must be set for each test positive and negative control, never can be omitted.

(4) before and after disinfection, sampling (positive control group and disinfection trial group), shall not be conducted in the same block.

(5) cotton swabs daub sampling are difficult to standardize, therefore should try to make the size of the cotton swabs, force uniform, absorb the quantity of sample liquid, washed bacteria was the weight of the hammer and so on successively.

(6) the sample must be timely detection. Room temperature storage must not exceed 2 h, otherwise should be 4 ℃ refrigerator, but also must not exceed 4 h.

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