Adenosine Triphosphate Bioluminescence Method Research Articles

Online monitoring system for qualitative and quantitative analysis of bioaerosols by combined ATP bioluminescence assay with loop-mediated isothermal amplification

Rapid detection of airborne pathogens is crucial in preventing respiratory infections and allergies. However, technologies aiming to real-time analysis of microorganisms in air remain limited due to the sparse and complex nature of bioaerosols. Here, we introduced an online bioaerosol monitoring system (OBMS) comprised of integrated units including a rotatable stainless-steel sintered filter-based sampler, a lysis unit for extracting adenosine triphosphate (ATP), and a single photon detector-based fluorescence unit. Through optimization of the ATP bioluminescence method and establishment of standard curves between relative luminescence units (RLUs) and ATP as well as microbial concentration, we achieved simultaneous detection of bioaerosols' concentration and activity. Testing OBMS with four bacterial and two fungal aerosols at a sampling flow rate of 10 to 50 L/min revealed an outstanding collection efficiency of 95 % at 30 L/min. A single OBMS measurement takes only 8 min (sampling: 5 min; lysis and detection: 3 min) with detection limits of 3 Pcs/ms photons (2.9 × 103 and 292 CFU/m3 for Staphylococcus aureus and Candida albicans aerosol). In both laboratory field tests, OBMS detected higher concentrations of bioaerosol compared to the traditional Andersen impactor and liquid biosampler. When combined OBMS with loop-mediated isothermal amplification (LAMP), the bioaerosol can be qualitative and quantitative analyzed within 40 min without the cumbersome procedures of sample pretreatment and DNA extraction. These results offer a high compressive and humidity resistance membrane filtration sampler and validate the potential of OBMS for online measurement of bioaerosol concentration and composition.

Can the adenosine triphosphate (ATP) bioluminescence assay be used as an indicator for hospital cleaning? - A pilot study.

In hospital cleaning, there is currently no standard for uniform monitoring of surface cleaning, either in Germany or internationally. One possibility for monitoring is the use of so-called objective methods for checking cleaning performance (e.g. fluorescence or adenosine triphosphate (ATP) method). The aim of the study was to monitor and evaluate the implementation of the adenosine triphosphate (ATP) bioluminescence assay as a cleaning indicator in everyday hospital cleaning, in order to verify its utility and effectiveness. In three phases, five frequently touched surfaces were examined with the ATP bioluminescence assay at different time points. 846 measurements were performed on the dermatology ward of a university hospital (phase 1), 1,350 measurements were performed on five different wards of the university hospital (phase 2), and 1,044 measurements were performed on five wards of another large hospital (phase 3). For this purpose, one structurally old and one structurally new ward as well as an intensive care unit (ICU), an outpatient clinic and a radiology department were selected for phases 2 and 3. With the ATP bioluminescence method, we were able to demonstrate a reduction in values after cleaning: before cleaning mean of ATP, 907 relative light units (RLU) (95% confidence interval [CI] 777; 1,038); after cleaning mean=286 RLU (CI=233; 495) (phase 1) and by intervention (five hours after daily cleaning mean=360 RLU (CI=303; 428); five hours after daily cleaning and two additional cleanings mean=128 RLU (CI=107; 152) (phase 3). The ATP values increased five hours after cleaning in phases 1 and 2, and eight hours after cleaning in phase 3. The structurally old wards had the highest ATP content, the ICU and the radiology department, among others, the lowest. In all phases, door handles showed both a reduction after cleaning or intervention and a subsequent increase in ATP values. Chair armrests, examination tables and door handles had high ATP values overall. The study shows ward differences both for cleaning effects and for the soiling characteristics of surfaces during the course of the day. In addition, it demonstrates the benefit of intermediate cleaning twice a day. It is noteworthy that structurally old stations and older inventory were more heavily soiled and, in some cases, more difficult to clean. The results show that the ATP bioluminescence method is suitable for detecting cleaning effects and can be used in everyday clinical practice for simple cleaning monitoring. Furthermore, it enables the detection of risk surfaces and easy-to-clean surfaces with significant re-soiling.

ATP bioluminescence method as a rapid tool for assessment of cleanliness of commercial animal transport trailers

Animal transportation is widely recognized as a significant risk for disease transmission. At present, cleanliness of animal transport trailers is mostly assessed through subjective visual inspection (i.e., ‘white-glove’ test), which may sometimes be supplemented by microbiological testing with results obtained after at least 2-3 days. In this study, the feasibility of using adenosine triphosphate (ATP) bioluminescence method as a rapid tool for objectively assessing animal transport trailer cleanliness was investigated. Eighteen newly-cleaned transport trailers were tested using both ATP bioluminescence and microbiological techniques. In each trailer, six (6) locations including floors, walls, ramps, partitions and trailer exterior surfaces were sampled using an ATP meter, and MacConkey and R2A agar contact plates. From a total of more than 500 paired samples collected from all the sampled trailers, significant correlation was found between ATP bioluminescence method and standard microbiological technique using R2A agar (r = 0.206; p = 0.001) and MacConkey agar (r = 0.154; p = 0.002) plates. Using a threshold or ‘Pass’ limit of 390 RLU per 100 cm2 of trailer surface, ATP method was able to provide a good objective measure of the surface cleanliness, thus may be considered as an additional tool available for rapid and less costly assessment of trailer surface cleanliness.

ATP MONITORING AS AN EXPRESS METHOD TO DETERMINE OBJECT CONTAMINATION

The article presents the results of indication of water and surface contamination. The researches were aimed at estimation of water safety and sanitary-hygienic condition of surfaces. To study the contamination of water samples and common surfaces, we used the traditional method and the bioluminescence-based express method. The bioluminescence method is based on determining the total amount of ATP (bacterial, somatic and extracellular) on contact surfaces and in water. The level of ATP (adenosine triphosphate) was determined using the Lumitester PD-30 (Kikkoman, Japan) luminometer device according to the manufacturer's instructions, using special test systems. The ATP bioluminescence method is commercially more available for simple and quick monitoring of the sanitary-hygienic control effectiveness according to the HACCP principles or international requirements. The traditional method of determining water (and other material) contamination was performed by inoculating water or surface wiping on a common nutrient medium and further cultivation under appropriate conditions. The strongest glowing reaction was observed in sea water, which can be explained by the presence of organic substances in it, while the bioluminescence values in potable (bottled) and filtered water tests were most close to control test. The results of testing the sanitary-hygienic condition of surfaces showed that the amount of adenosine triphosphate exceeded the limits in almost all tested objects. However, a slight exceeding of adenosine triphosphate level was observed on the internal surface of new (plastic) food containers. The investigations performed show that the bioluminescence-based express method can be used as a primary control that gives immediate information about the contamination of both surfaces and liquids. Using the bioluminescence method can shorten the time of the study and therefore reduce the cost of the test. However, when determination of qualitative and quantitative composition of the tested object’s microbiota is necessary, then the classical microbiological control must be performed.

Assessment of percent positive agreement between fluorescent marker and ATPase for environmental cleaning monitoring during sequential application in an intensive care unit

Terminal room cleaning is critical in preventing pathogen transmission; however, the optimal cleaning effectiveness assessment modality is still being investigated. We sequentially compared cleanliness assessment agreement between a fluorescent marker and an adenosine triphosphate bioluminescence method, finding no significant differences between modalities.

The Contamination Degree of Characteristic Fungi in Jet Fuel Detected by ATP Bioluminescence

Adenosine Triphosphate (ATP) lysate of the characteristic fungi in jet fuel was screened and three fluorescein-luciferase systems were compared in the paper. The correlation between ATP bioluminescence and traditional plate count for measuring the number of microorganism was also investigated and then a method of detecting the contamination degree of the characteristic fungi of jet fuel by ATP bioluminescence were initially established. The results show that the effect of ATP extraction taking surfactant Benzyldimethylhexadecylammonium chloride (BAC) as the microbial lysate was optimal and the optimal concentration and the action time were 0.15% and 30s respectively; The fluorescein-luciferase system after screening has a good detection limit, up to 10-15mol ATP; using ATP bioluminescence and traditional plate method to measure the microbial quantity, they have good correlation. Utilizing ATP bioluminescence to detect the contamination degree of the characteristic fungi in jet fuel can shorten the test time to 10min, which is suitable for rapid detection and has good application prospects.

Preventing Healthcare-Associated Infections by Monitoring the Cleanliness of Medical Devices and Other Critical Points in a Sterilization Service.

It is well known that the common goal of all central sterile supply departments (CSSDs) is to prevent healthcare-associated infections. Such infections entail high costs to society, not only economic but also social. Therefore, delivering safe medical devices and guaranteeing a positive contribution to the control of healthcare-associated infections form the main responsibilities of a CSSD. The monitoring of the effectiveness of medical device cleaning processes is highly recommended. However, ensuring a flawless environment for the preparation, assembly, and packaging of medical devices and clean handling of sterilized items is crucial to achieving the goal of safe medical devices. This study analyzed not only the cleanliness of surgical instruments but also two critical aspects of the surrounding environment: the cleanliness of work surfaces and the cleanliness of workers' hands. To evaluate the cleanliness of surgical instruments, two methods were used: the adenosine triphosphate (ATP) detection method and a residual protein test. It was not the intention of this work to make an exhaustive comparison of these methods. The ATP bioluminescence method was also used for monitoring the cleanliness of work surfaces and workers' hands. The aims of this study were to establish the most suitable method of evaluating the cleanliness of reusable medical devices in the CSSD and to assess the quality of the environment. Assessing the surgical instruments, work surfaces, and staff hands for cleanliness allowed the identification of possible contamination sources and to correct them by improving cleaning/disinfection protocols. Furthermore, the use of ATP monitoring tests of workers' hands highlighted the importance of staff compliance with good practice guidelines. Thus, these results have a positive impact on the CSSD quality system and, consequently, on patient safety.

Fast monitoring of indoor bioaerosol concentrations with ATP bioluminescence assay using an electrostatic rod-type sampler.

A culture-based colony counting method is the most widely used analytical technique for monitoring bioaerosols in both indoor and outdoor environments. However, this method requires several days for colony formation. In this study, our goal was fast monitoring (Sampling: 3 min, Detection: < 1 min) of indoor bioaerosol concentrations with ATP bioluminescence assay using a bioaerosol sampler. For this purpose, a novel hand-held electrostatic rod-type sampler (110 mm wide, 115 mm long, and 200 mm tall) was developed and used with a commercial luminometer, which employs the Adenosine triphosphate (ATP) bioluminescence method. The sampler consisted of a wire-rod type charger and a cylindrical collector, and was operated with an applied voltage of 4.5 kV and a sampling flow rate of 150.7 lpm. Its performance was tested using Staphylococcus epidermidis which was aerosolized with an atomizer. Bioaerosol concentrations were measured using ATP bioluminescence method with our sampler and compared with the culture-based method using Andersen cascade impactor under controlled laboratory conditions. Indoor bioaerosol concentrations were also measured using both methods in various indoor environments. A linear correlation was obtained between both methods in lab-tests and field-tests. Our proposed sampler with ATP bioluminescence method may be effective for fast monitoring of indoor bioaerosol concentrations.

Validation of ATP bioluminescence as a tool to assess antimicrobial effects of mouthrinses in an in vitro subgingival-biofilm model

Objectives. The aim of this investigation was to evaluate whether the adenosine triphosphate (ATP) bioluminescence method is an appropriate tool to assess the efficacy of antiseptic mouthrinses in terms of quantitative reductions of total viable microbial counts in mixed biofilm populations in vitro. Study Design. Three mouthrinses, containing respectively, chlorhexidine and cetylpyridinium chloride (CHX/CPC), essential oils (EO) and amine fluoride/stannous fluoride (AFSF), as well as Phosphate Buffered Saline (PBS) used as control, were tested in an in vitro static biofilm model by ATP bioluminescence and compared to culture method. Biofilms were grown on saliva-coated hydroxyapatite disks for 72 hours and then exposed for 1 minute to the mouthrinse or control by immersion. The antibacterial effect of the rinses was tested by analysis of variance. The reliability of the ATP bioluminescence method was assessed by calculating the Pearson correlation coefficients when compared to the viable cell counts obtained by culture. Results. Using ATP bioluminescence, the antimicrobial activity of the tested mouthrinses was demonstrated when compared to the PBS control. The ATP bioluminescence values were significantly correlated (0.769, p<0.001) to the viable cell counts. CHX/CPC and AFSF showed similar antimicrobial activity, although AFSF had a less homogeneous effect, being both more effective than the EO rinse. Conclusion. ATP bioluminescence viability testing may be considered a useful tool to assess the in vitro efficacy of antibacterial compounds. In the proposed model, CHX/CPC and AFSF containing mouthrinses demonstrated superior antimicrobial activity, as compared to EO rinses, in a multispecies biofilm model. Key words:Biofilm, ATP bioluminescence,mouthrinse, essential oils, chlorhexidine, amine fluoride/stannous fluoride.

ATP bioluminescence rapid detection of total viable count in soy sauce

The adenosine triphosphate (ATP) bioluminescence rapid determination method may be useful for enumerating the total viable count (TVC) in soy sauce, as it has been previously used in food and beverages for sanitation with good precision. However, many factors interfere with the correlation between total aerobic plate counts and ATP bioluminescence. This study investigated these interfering factors, including ingredients of soy sauce and bacteria at different physiological stages. Using the ATP bioluminescence method, TVC was obtained within 4 h, compared to 48 h required for the conventional aerobic plate count (APC) method. Our results also indicated a high correlation coefficient (r = 0.90) between total aerobic plate counts and ATP bioluminescence after filtration and resuscitation with special medium. The limit of quantification of the novel detection method is 100 CFU/mL; there is a good linear correlation between the bioluminescence intensity and TVC in soy sauce in the range 1 × 10(2) -3 × 10(4) CFU/mL and even wider. The method employed a luminescence recorder (Tristar LB-941) and 96-well plates and could analyse 50-100 samples simultaneously at low cost. In this study, we evaluated and eliminated the interfering factors and made the ATP bioluminescence rapid method available for enumerating TVC in soy sauce.

Application of ATP bioluminescence method to characterize performance of bioaerosol sampling devices

In this study we developed and tested an adenosine triphosphate (ATP) bioluminescence-based protocol for rapid characterization of bioaerosol sampling devices when collecting bacterial aerosols. Initial calibration of the method was performed using commercially available ATP assays to quantify known bacterial biomass in liquid samples. Here, the relative luminescence units (RLUs) indicative of total ATP content was plotted against total bacterial counts obtained using acridine orange direct count epifluorescence microscopy. The developed calibration curves were used to evaluate the collection performance of a BioSampler (SKC Inc., Eighty Four, PA) when sampling Pseudomonas fluorescens and Bacillus subtilis bacteria at three different sampling flow rates. The average collection efficiency for P. fluorescens and B. subtilis over all flow rates was 69% and 54%, respectively. Collection efficiencies determined by the ATP method for both bacteria at all flow rates agreed well (within 5%) with the acridine orange direct counts. Further, the ATP-based method allowed insights into the losses and fate of particles inside the sampling device. It was determined that a majority of losses occur in sampler's inlet. Such evaluation of losses is very difficult to perform using microscopy or culture-based techniques. Thus, the ATP-based technique could be reliably used as a tool for rapid and detailed evaluation of bioaerosol sampling devices.

Combining biofunctional magnetic nanoparticles and ATP bioluminescence for rapid detection of Escherichia coli

A rapid, specific and sensitive method for assay of Escherichia coli ( E. coli) using biofunctional magnetic nanoparticles (BMNPs) in combination with adenosine triphosphate (ATP) bioluminescence was proposed. The BMNPs were fabricated by immobilizing a specific anti- E. coli antibody on the surface of amine-functionalized magnetic nanoparticles (about 20 nm in diameter), and then was applied to capture the target bacteria E. coli from samples. The BMNPs exhibited high capture efficiency to E. coli. Transmission electron microscope (TEM) images showed that the BMNPs were bound to the surface of entire E. coli cells. The target bacteria became magnetic so that could be isolated easily from the sample solution by employing an external magnetic field. The concentration of E. coli captured by the BMNPs was then detected by an ATP bioluminescence method. The optimization of ATP measurement was carried out to improve the detection sensitivity. The proposed method was applied to detect the E. coli inoculated into pasteurized milk with low detection limit (20 cfu/mL) and short detection time (about 1 h).

The detection of food soils and cells on stainless steel using industrial methods: UV illumination and ATP bioluminescence

Open food contact surfaces were subjected to organic soiling to provide a source for transfer of microbial cells. Rapid industrial methods used for the detection of residual cells and soil e.g. ATP (adenosine triphosphate) bioluminescence and an ultraviolet (UV) light detection method were assessed for their ability to detect organic soils, or organic soil–cell mix on surfaces. A range of soils (complex [meat extract, fish extract, cottage cheese extract]; oils [cholesterol, fish oil, mixed fatty acids]; proteins [bovine serum albumin, fish peptones casein]; carbohydrates [glycogen, starch, lactose]); was used. Under UV, oily soils, mixed fatty acids, cholesterol and casein were detected at low concentrations, with detection levels ranging from 1% to 0.001% for different substances. Glycogen was the most difficult substance to detect at lower concentrations. Using UV wavelength bands ( λ) of 330–380 nm, 510–560 nm and 590–650 nm, wavelength bands of 330–380 nm, illuminated most of the soils well, whilst the wavelength band of 510–560 nm illuminated the fish extract, cholesterol and fatty acids; the 590–650 nm wavelength band illuminated the lactose. Soils at all concentrations were detected by the ATP bioluminescence method; the complex soils gave the highest readings. When complex soils were combined with Listeria monocytogenes Scott A or a non-pathogenic Escherichia coli O157:H7, ATP measurements increased by 1–2 logs . For UV illumination, the L. monocytogenes and cheese combination was the most intensely illuminated, with E. coli and meat the least. UV illumination is a simple well established method for detecting food soil, with little change in findings when microorganisms are included. Performance can be enhanced in certain circumstances by altering the wavelength. ATP bioluminescence is a proven system for hygienic assessment being especially useful in the presence of microorganisms rather than organic soil alone.

Adenosine Triphosphate Bioluminescence as a Method to Determine Microbial Levels in Scald and Chill Tanks at a Poultry Abattoir

According to Hazard Analysis of Critical Control Points (HACCP) programs developed for the poultry industry, poultry processing waters should be actively monitored to minimize cross-contamination between chicken carcasses. In order to monitor HACCP programs, a test is required that provides results on a real time basis. A modified adenosine triphosphate (ATP) bioluminescence test has been developed that can assess microbial levels in poultry processing waters within 15 min. A study was conducted to determine the effectiveness of this test for examining scald, prechill, and chill tank waters. The results showed that the modified ATP bioluminescence method gave results comparable to plate counts. The microbial levels were dependent on the tank and the time of sampling. The highest microbial levels were detected in the scald tank. In all three tanks, the microbial levels increased over time during the day.

Adenosine Triphosphate Bioluminescence for Hygiene Monitoring in Health Care Institutions

An investigation was conducted to assess the practical use of an adenosine triphosphate (ATP) bioluminescence assay to evaluate the effectiveness of cleaning and sanitizing meat slicers in eight health care institutions. The ATP bioluminescence assay was compared to conventional swabbing techniques using standard plate count to enumerate microbial load. Assays were performed on meat slicers before use, after slicing a meat product and after sanitizing. There was a general overall agreement in results obtained by both methods but the ATP assay gave a better indication of the cleanliness of the meat slicer as it was able to detect the presence of meat residues left on the blade after improper sanitation. Results were available within 5 min using the ATP bioluminescence method, thus providing an opportunity for immediate remedial action.