Rapid Microbiological Methods Research Articles

A metagenomics-based workflow for the detection and genomic characterization of GBS in raw freshwater fish.

The need for a rapid and accurate food microbiological testing method is apparent for a timely and effective foodborne outbreak response. This is particularly relevant for emerging foodborne pathogens such as Group B Streptococcus (GBS) whose associated food safety risk might be undercharacterized. By using GBS in raw freshwater fish as a case example, this study describes the development of a metagenomics-based workflow for rapid food microbiological safety testing and surveillance. This study can inform as a working model for various foodborne pathogens in other complex food matrices, paving the way for future methodological development of metagenomics for food microbiological safety testing.

Genital ulcer disease: Study of factors associated and utility of rapid diagnostic methods in current pandemic.

Genital ulcer disease (GUD) is regarded as the most difficult conundrum for clinicians, as valid and rapid methods to diagnose them are limited, especially in India. The sexual behavior of an individual plays a role in the causation of GUD, and it depends on psychological, behavioral, and social well-being, which was significantly affected during the COVID-19 pandemic. Hence, the present study was undertaken to determine the etiology of genital ulcers by using simple, rapid microbiological methods and to study various factors associated with them. Various rapid tests like staining (Giemsa, Gram, and Diff Quick) and serology for antibody detection were used to diagnose agents causing genital ulcers among 71 suspected patients during the study period of 1 year. After due informed consent, sociodemographic and behavioral factors were recorded in proforma and analyzed. Rapid tests could aid in diagnosing 47.9% of cases of GUD. Turnaround time was less than 60 minutes for all tests carried out. Genital herpes was the most common GUD followed by syphilis. Male gender (74.6%), age group of more than 34 years old (62%), literate (71.8%), and earning (70.4%) constituted most of the population presenting with GUD. Behavioral factors like promiscuity with a known person (83.1%) had a significant association (P 0.04) with GUD. Rapid diagnostic tests could assist in the early treatment of GUD cases. Factors like literacy, financial independence, and promiscuity with known individuals were associated with GUD in our study during the COVID-19 pandemic.

The Challenges of The Diagnostic and Therapeutic Approach of Patients with Infectious Pathology in Emergency Medicine.

A retrospective observational study was performed on 657 patients, from whom clinical samples (urine, purulent secretions, blood cultures, etc.) were collected for microbiological diagnosis in the first 3 days after presentation in the ED. The identification of pathogens and the antimicrobial susceptibility testing with minimum inhibitory concentration determination were carried out according to the laboratory protocols. From the 767 biological samples analyzed, 903 microbial isolates were identified. E. coli was most frequently isolated (24.25%), followed by Klebsiella spp., S. aureus (SA), and non-fermentative Gram-negative bacilli. E. coli strains maintained their natural susceptibility to most antibiotics tested. In the case of Pseudomonas spp. and Acinetobacter spp., increased rates of AMR were identified. Also, 32.3% of SA strains were community-acquired MRSA. The introduction of rapid microbiological diagnostic methods in emergency medicine is imperative in order to timely identify AMR strains and improve therapeutic protocols.

Bloodstream Infections: Comparison of Diagnostic Methods and Therapeutic Consequences between a Hospital in a Resource-Limited Setting and Two French Hospitals.

In recent years, the diagnosis of bloodstream infections has been complemented by rapid microbiological methods, unattainable to most clinical laboratories in resource-limited settings. We evaluated the impact of their shortage on antibiotic therapy adequacy. We conducted a prospective multicenter cohort study including 150 adult Gram-negative bacilli bacteremia episodes, evenly distributed across three university hospitals: one in Lebanon, a resource-limited setting, and two in France, a resource-rich setting. Previous colonization by multidrug-resistant organisms (MDRO) was significantly more prevalent among the Lebanese than the French group of patients (16/50 vs. 5/100; p < 0.01). Bloodstream infections by carbapenemase-producing Enterobacterales and other MDRO were higher among the Lebanese than the French group of patients (25/50 vs. 12/100; p < 0.01). For the French group, rapid identification of species and mechanisms of resistance significantly shortened turnaround time for definitive laboratory diagnosis and increased antibiotic therapy adequacy. No statistically significant differences were noted in targeted antibiotic therapy between the two groups. This study suggests that, in settings where bacterial resistance is prevalent, rapid microbiological methods have not provided any additional value. The clinical and economic impact of rapid microbiological methods will likely depend on local CPE, VRE, and other MDRO epidemiology and are areas for future research.

SYNERGIZING FOOD SAFETY, QUALITY AND GENETIC IMPROVEMENT: THE INTERSECTION OF FOOD MICROBIOLOGY AND PROCESSING

The fusion of food microbiology and processing has given rise, to a field that encompasses food safety, quality and genetic enhancement. This convergence plays a role in ensuring the safety, nutritional value and overall superiority of the food we consume. By integrating knowledge of microbiology with genetic improvement strategies significant progress has been made in preservation methods and food processing techniques. Innovative processing technologies have proven effective in mitigating the presence of microorganisms and extending the shelf life of food products thereby enhancing food safety. Techniques such as high pressure processing pulsed fields and ultraviolet light have gained recognition for their ability to regulate microbial populations and maintain the nutritional integrity of food. This comprehensive approach to food safety combines principles from both microbiology and processing to address challenges and safeguard health. Concurrently strategies in the realm of food microbiology and processing strive to enhance the quality and nutritional value of our meals. Probiotics, which are microorganisms that offer health benefits are employed to promote wellbeing, bolster function and overall vitality. Through engineering probiotics can be customized to target health advantages. The presence of antibiotic microorganisms like Listeria monocytogenes and Salmonella poses a risk, to food safety. To tackle this problem various approaches are utilized, such as enhancement, focused antimicrobial techniques and rigorous compliance, with safety protocols. The utilization of rapid microbiological methods, such as PCR and metagenomics, enables improved monitoring and detection of evolving microorganisms, thereby enhancing food safety. DNA-based techniques are used to combat microbial food fraud, authenticate products, detect adulteration, and trace the origins of ingredients, all of which contribute to consumer trust and the integrity of the food supply chain.

Validation of a rapid microbiological method for the detection and quantification of the Burkholderia cepacia complex in an antacid oral suspension.

Despite the broad adoption of Soleris® technology in the food industry as semiquantitative method, they are almost completely unexplored in the pharmaceutical industry as a quantitative method for quantification of Burkholderia cepacia complex. The efficacy of an automated growth-based system for a quantitative determination of the Burkholderia cepacia complex in an antacid oral suspension was studied. The main purpose of this validation study was to prove that the alternative method's entire performance is not inferior to the conventional method for a quantitative determination of Burkholderia cepacia complex. The antacid oral suspension's preservatives were neutralized, leading to the Burkholderia complex's recovery by means of the alternative method and the reference method. A calibration curve was generated for each strain by plotting DTs relative to the corresponding log CFU values. An equivalence of results was done through the construction of calibration curves that allowed the establishment of numerically equivalent results between the enumeration data from the reference method and the alternative method. Thus following the guidelines of USP, essential validation parameters were shown, such as equivalence of results (CC > 0.95), linearity (R2 >0.9025), accuracy (% recovery >70%), operating range, precision and ruggedness (DS < 5 and CV < 35%), specificity (inclusivity and exclusivity), limit of detection, and limit of quantification. It was shown that all the test results obtained from the alternative method were in statistical agreement with the standard method. Thus this new technology was found to meet all the validation criteria needed to be considered to be an alternative method for the quantification of the Burkholderia complex in the antacid oral suspension tested. mplementation of alternative methods can offer benefits in execution and automation while improving accuracy, sensitivity, and precision and reduce the microbiological process time compared to the traditional ones.

Molecular Characterization of Microbial Quality of Ready-to-eat Salads using Multi-locus Sequence Typing

Uncertainty persists concerning the role of ready-to-eat (RTE) salad as a bacterial reservoir. The attention paid to food safety by international agencies and international regulations has not improved food safety despite technological advancements, RTE salad’s microbiological quality and safety still seems challenging. The present study’s objective was to detect any microorganism in the RTE salads sold in supermarkets of Riyadh, Saudi Arabia, and to interpret the susceptibility pattern of isolated bacteria to a set of antimicrobials. Phenotypic methods and biochemical analysis were used to identify the isolated bacterium from each salad sample. Antibiograms of the isolated bacteria was determined by VITEK system 2. Multi-locus sequence typing (MLST) was performed for 15 Escherichia coli isolates for investigating evolutionary relationship and genetic analysis. The culture-based technique showed that the major species identified in samples were Aeromonas spp., Acinetobacter spp., E. coli, Roultella ornithinolytica, Citrobacter koseri, Luciferciaadec arboxylata, Klebsiella oxytoca, and Aerococcus viridians. Remarkably, Acinetobacter spp. showed the highest antibiotic resistance to erythromycin, nitrofurantoin and co-trimoxazole. ST 1887 was the most common one traced in 3 E. coli isolates, when total of 12 STs (sequence types) were specified to 15 isolates. A total of three clonal complexes (CC); CC-12, CC-14 &amp; CC-23 were reported in this study. Implementing an accurate, rapid, and easy microbiological analysis method could be valuable for providing higher quality products. Based on the obtained results, dedicated regular RTE salad quality monitoring is recommended, and hand hygiene should be maintained while handling and packaging of RTE.

Improvement of the Pour Plate Method by Separate Sterilization of Agar and Other Medium Components and Reduction of the Agar Concentration.

Although the pour plate method is widely employed in microbiological quality control, it has certain drawbacks, including having to melt the culture medium before seeding. In this study, the preparation of the culture medium was modified by using a lower concentration of agar (10 g/L), which was separated from the nutrients during sterilization. The new protocol was assessed in media frequently used in microbiological quality control of food, cosmetics, and pharmaceutical products, with tryptic soy agar (TSA), Sabouraud 4% dextrose agar (SDA), and violet red bile glucose agar (VRBG). In comparison with the conventionally produced media, the modifications significantly improved the growth of Saccharomyces cerevisiae in SDA, Staphylococcus aureus, Salmonella enterica subsp. enterica serovar Typhimurium, and Candida albicans in TSA and Escherichia coli ATCC 8739 and ATCC 25922 and S. Typhimurium in VRBG. The modified VRBG was also more selective for Pseudomonas aeruginosa. Regarding physicochemical properties, a significantly lower pH was observed in TSA and VRBG and lower strength values in TSA. Sterilizing agar separately from the other components of the medium and reducing the agar concentration to 10 g/L can improve microorganism growth and enhance the selectivity of differential media in the pour plate method. These modifications could facilitate the automation of this culture technique. IMPORTANCE In the era of rapid microbiological methods, there is a need to improve long-established culture techniques. Drawbacks of the pour plate method include having to melt each medium separately before seeding. For this technique, we demonstrate that separating the agar from the other components of commonly used media during sterilization and reducing the agar concentration to 10 g/L can enhance microbial growth. The new protocol could have advantages in routine laboratory practice because less agar is required and the same molten agar suspension can be used to prepare different media. Moreover, these modifications could facilitate the automation of the pour plate method.

Performance equivalence and validation of a rapid microbiological method for detection and quantification of yeast and mold in an antacid oral suspension.

Alternative and rapid microbiological methods are effective and profitable technologies that can be safely use in the pharmaceutical industry. This report studies the efficacy of the Soleris® direct yeast and mold automated method for the quantitative detection of C. albicans and A. brasiliensis at three different microbial bioburden. Validation testing was carried out using an antacid oral suspension (aluminum hydroxide 4% + magnesium hydroxide 4% + simethicone 0.4%). Equivalence of data between detection time vs. colony-forming units was established for both, alternative and conventional methodologies. Using probability of detection, linear Poisson regression, Fisher's test, and multifactorial analysis of variance ANOVA, all results from the rapid method were prove to be in statistical agreement with the reference plating procedures. The limits of detection and quantification were statistically similar for both methods (Fisher's exact test, P >0.05), showing that the alternative method was not inferior in performance to the reference method. Essential validation parameters such as precision (standard deviation < 5, coefficient of variance < 35%), accuracy (>70%), linearity (R2 >0.9025), and ruggedness (ANOVA, P <0.05) were determined. It was proved that all test results obtained from the alternative method were in statistical agreement with the standard plate count method. Thus, this new technology was found to meet all the validation criteria needed to be considered as an alternative method for yeast and mold quantification in the antacid oral suspension tested.

Challenges Encountered in the Implementation of Bio-Fluorescent Particle Counting Systems as a Routine Microbial Monitoring Tool.

The transition from traditional growth-based microbial detection methods to continuous bio-fluorescent particle counting methods represents a paradigm shift, because the results will be non-equivalent in terms of microbial counts, and a continuous, rather than periodic, data stream will be available. Bio-fluorescent particle counting technology, a type of rapid microbiological method, uses the detection of the intrinsic fluorescence of microbial cells to enumerate bioburden levels in air or water samples, continuously. The reported unit is commonly referred to as an autofluorescence unit, which is not dependent upon growth, as is the traditional method. The following article discusses challenges encountered when implementing this modern technology, and the perspective from a consortium of four industry working groups on navigating these challenges.

The Value of Metagenomic Next-Generation Sequencing in Hematological Malignancy Patients with Febrile Neutropenia After Empiric Antibiotic Treatment Failure.

BackgroundIt was crucial to use empirical antibiotics in febrile neutropenia (FN) patients. However, most patients still died from infection due to poor efficacy. Metagenomic next-generation sequencing (mNGS) is a rapid microbiological diagnostic method. The value of mNGS in patients with FN remains to be studied, especially after empiric antibiotic treatment.MethodsWe retrospectively analyzed the differences between mNGS and the traditional methods in 192 patients with hematological malignancies who have received empiric antibiotic treatment. Samples were collected when patient had chills or half an hour before peak body temperature. And we compared the differences between FN and non-FN patients, mainly including types of pathogens and the diagnostic value of different pathogens.ResultsDespite receiving empirical treatment, the pathogen detection rate of mNGS was significantly higher than the traditional method (80.21% vs 25.00%, P<0.001). And it has obvious advantages in detecting mixed pathogens infection (80.21% vs 4.17%, P<0.001). Then, we found that mNGS saw more pathogens in the FN than in the non-FN group, especially fungus. 21/33 (63.63%) of FN patients was diagnosed with fungal infections. The fungal detection rate in FN was significantly higher than non-FN group (32.35% vs 12.22%, P=0.001). Besides, the sensitivity of mNGS was higher than the traditional methods in both FN and non-FN group (P<0.001), but no significant difference in specificity (P>0.05). In the FN group, empiric antibiotic treatment of 46/102 (45.10%) patients did not treat all the pathogens detected by mNGS. After adjusting the antimicrobial regimen according to the results of mNGS, the effective rate at 72 hours and 7 days was 22/46 (47.83%) and 24/102 (52.17%), respectively.ConclusionmNGS had a significant impact on the diagnosis of infection and the second-line antimicrobial therapy in FN. mNGS plays a more important role in FN patients, especially in the diagnosis of fungal infections.PurposeFirstly, we compared the difference between mNGS and the traditional methods in the diagnosis of infection. Secondly, we assessed the value of mNGS in FN patients by comparing it with non-FN patients, including types of pathogens and the diagnostic value of different pathogens. In order to show that mNGS plays a more important role in FN.

Serum Calprotectin Levels in Dogs with Diarrhea

Background: Diarrhea induced by infectious factors may lead to significant health problems in dogs. Canine parvovirus (CPV), canine coronavirus (CCV), canine distemper virus (CDV), Giardia spp., Escherichia coli (E. coli), and Salmonella spp. are the important infectious agents that may induce diarrhea in dogs. The present study aimed to investigate the effect of CPV, CCV, CDV, Giardia spp., E. coli, and Salmonella spp. infections on the change in serum calprotectin (Calp) concentration.Materials, Methods &amp; Results: A total of 30 dogs were enrolled in the study. The study dogs were divided into 3 groups. Healthy animals as confirmed by clinical examination and animals negative for the specified pathogens were placed in Group 1. Animals infected by one or more agents, including CPV, CCV, CDV, and Giardia spp., but negative for E. coli or Salmonella spp. were placed in Group 2. Finally, animals positive for E. coli or Salmonella spp. and infected or not infected by one or more agents, including CPV, CCV, CDV, and Giardia spp., were placed in Group 3. Stool samples and rectal and conjunctival swab samples were collected to investigate the etiologic agents that induced diarrhea. Blood samples were collected through vena cephalica antebrachii for hematological and biochemical examinations. The samples were obtained via routine clinical examinations at the Prof. Dr. Servet SEKİN outpatient clinic at Dicle University Veterinary Faculty. CPV, CCV, CDV, and Giardia spp. diagnoses were made based on immunochromatographic test kits. The bacteriological analysis of stool samples were used to diagnose E. coli and Salmonella spp. infection. Serum Calp concentrations were measured by Enzyme-Linked Immunosorbent Assay (ELISA). The analysis of swab and stool samples by immunochromatographic rapid diagnosis kits and microbiological methods showed that 5 animals were infected with CPV, 10 with CCV, 6 with CDV, 3 with Giardia spp., 12 with E. coli, and 2 with none of the specified agents. Total leukocyte count (WBC), lymphocyte (Lym) (%), and granulocyte (Gra) (%) values were higher in the diarrheal dogs when compared with the control group. In the biochemical examination of serum samples, TBIL and P levels were higher and Na levels were lower in Group 3 when compared to the control group (P = 0.025, 0.024, and 0.018, respectively). TP and Alb values were lower in Group 2 compared to Groups 1 and 3 (P = 0.001 and 0.019 for TP, P = 0.000 and 0.001 for Alb, respectively). There was a statistically significant difference in CK levels between Group 1 and Group 2 (P = 0.013). Serum Calp level was higher in the E. coli infected group (Group 3) compared to the other groups, no significant differences were noted between the groups (P &gt; 0.05).Discussion: In conclusion, to the best of authors knowledge, this study is the first to evaluate serum Calp levels in dogs with diarrhea induced by viral, bacterial, and protozoan infections. The Calp level was higher in the sick dogs that were infected by at least one agent, including CPV, CCV, CDV, and Giardia spp., and were at the same time E. coli positive when compared with the control group and the group without E. coli infections. It was concluded that new studies could be useful to reveal the diagnostic importance of serum Calp concentration in dogs with diarrhea and that these results may contribute to future studies in this area.Keywords: calprotectin, diarrhea, dog, serum.

Combating QC challenges and special needs of cell and gene therapy production

<div class="authors authors-lg"><div class="author"><div class="author-img"><img src="https://cdn.insights.bio/uploads/Félix Montero copy.JPG" data-image="1"></div><div class="author-det"> <b>Félix Montero-Julian</b> is the Scientific Director for the Healthcare Business of bioMérieux (Lyon, France). Felix has over 25 years of experience in industrial and clinical diagnostics, and previously served as the Chemunex Ivry-sur-Seine site &amp; R&amp;D Director in bioMérieux. Felix is a member of different scientific organizations, such as the Parenteral Drug Association (MD, USA), and served as an expert in a panel for the Development of Compendial Rapid Sterility Tests for the US Pharmacopeia (MD, USA). Félix has been, and continues to be, extensively involved in the implementation and acceptance of rapid and alternative microbiological methods. He is a prominent speaker at congresses and conferences and a regular contributor to bioMérieux scientific white papers. Félix has extensive technical experience that includes the development of in vitro diagnostic and research use reagents and applications, cell and tissue culture systems, alternative and rapid microbiological methods, sterility testing, Mycoplasma detection, compendia methods, and cell and gene therapy processes. </div></div></div><div class="authors authors-lg"><div class="author"><div class="author-img"><img src="https://cdn.insights.bio/uploads/Rey Mali copy.png" data-image="1"></div><div class="author-det"> <b>Rey Mali </b>joined Accellix as its Vice President Sales and Marketing on July 2019. She has served as a Product Development and Marketing leader for the prior 14 years in both hi-tech and biotech companies. Rey has a graduate degree in Medical Research and a MBA from The University of California, Berkeley and started her career in pharmaceutical sales at Merck Pharmaceuticals. She then joined a biotech startup that was acquired by Life Technologies (Thermo Fisher) as a Field Applications Scientist (FAS) increasing customer acquisition and retention and later was promoted to a Global Product Manager role at Life Technologies, where she launched and managed products in the molecular biology space. She then transitioned to a Sr. Product Marketing Manager at Illumina, where she launched and managed products in the sequencing space, and later served as the Product Marketing Director at Twist Bioscience leading the global marketing team towards commercial product launches and a successful IPO. Recently, she has been working at the Chan Zuckerberg Initiative (CZI) leading the Product Marketing efforts of free technological tools to advance medical research. </div></div></div>

A Study Into the Use of Milliflex Quantum as Rapid Microbial Detection Method for Microorganisms in Pharmaceutical...

Control of pharmaceutical water systems represents an essential part of Good Manufacturing Practice and embedded within this is the quality control testing of water systems for viable microorganisms, and subjecting the data obtained to trend analysis. This has traditionally been achieved by membrane filtration and the use of a culture medium. While such test methodologies can recover a level of the bioburden present, the incubation times are lengthy. To address the problem of time-to-result, rapid microbiological methods offer an alternative approach. One such example is the Milliflex Quantum, which is the focus of this article.

Loop-mediated isothermal amplification combined with immunomagnetic separation and propidium monoazide for the specific detection of viable Listeria monocytogenes in milk products, with an internal amplification control

Nowadays, the most widely accepted rapid methods in food microbiology rely on nucleic acid amplification such as PCR/real-time PCR. A major claimed limitation of these methods is their incapacity to differentiate among viable and non-viable microorganisms. In the present study we report the development of a novel multiplex loop-mediated isothermal amplification method which, by combining immunomagnetic separation, to concentrate and purify the bacteria, along with propidium monoazide to block the amplification of DNA from non-viable microorganisms. The method allowed to specifically detect viable Listeria monocytogenes present in milk products. We designed an internal amplification control to rule out false negative results due to reaction inhibition, which is differentiated from L. monocytogenes by a simple melt curve analysis. Overall, the methodology provided results higher than 95% in terms of sensitivity, specificity and accuracy, as well as a Cohen's k of 0.97, reaching a limit of detection of 2.7 cfu/25 g. In samples inoculated with up to 106-107 cfu of dead microorganisms, the method demonstrated capable of effectively eliminating undesired amplification.

Use of 27G needles improves sensitivity and performance of ATCC anaerobe reference microorganism detection in BacT/Alert system

Effective detection of microbiological contaminations present in medicinal cellular products is a crucial step to ensure patients’ safety. In recent decades, several rapid microbiological methods have been developed and validated, but variabilities linked to the use of different resources have led to discordant validation of methods and performance results. Considering this, while developing an in-house BacT/Alert-based method, we evaluated all of the materials used in its validation. Of particular importance, we noticed that the syringe gauge used to inject the samples into the bottles was crucial to obtain robust results. We chose to conduct a comparative test between the BacT/Alert system and the compendial method described in the European Pharmacopoeia, using five dilutions of nine reference microorganism strains and 21G or 27G needles. Our results confirmed that the BacT/Alert system is a valid and faster alternative method to assess sterility of clinical cell therapy products, and that the use of 27G needles increases its sensitivity to detect reference anaerobe microorganisms.

Rapid identification of microbial contaminants in pharmaceutical products using a PCA/LDA-based FTIR-ATR method

Microbiological quality of pharmaceuticals is fundamental in ensuring efficacy and safety of medicines. Conventional methods for microbial identification in non-sterile drugs are widely used; however they can be time-consuming and laborious. The aim of this paper was to develop a chemometric-based rapid microbiological method (RMM) for identifying contaminants in pharmaceutical products using Fourier transform infrared with attenuated total reflectance spectrometry (FTIR-ATR). Principal components analysis (PCA) and linear discriminant analysis (LDA) were used to obtain a predictive model capable of distinguishing Bacillus subtilis (ATCC 6633), Candida albicans (ATCC 10231), Enterococcus faecium (ATCC 8459), Escherichia coli (ATCC 8739), Micrococcus luteus (ATCC 10240), Pseudomonas aeruginosa (ATCC 9027), Salmonella typhimurium (ATCC 14028), Staphylococcus aureus (ATCC 6538), and Staphylococcus epidermidis (ATCC 12228) microbial growth. FTIR-ATR spectra provide data on proteins, DNA/RNA, lipids, and carbohydrates constitution of microbial growth. Microbial identification provided by PCA/LDA based on FTIR-ATR method were compatible with those obtained using traditional microbiological methods. The chemometric-based FTIR-ATR method for rapid identification of microbial contaminants in pharmaceutical products was validated by assessing the sensitivity (93.5%), specificity (83.3%), and limit of detection (17-23 CFU/mL of sample). Therefore, we propose that FTIR-ATR spectroscopy may be used for rapid identification of microbial contaminants in pharmaceutical products and taking into account the samples studied

A Discussion on Bio-Fluorescent Particle Counters: Summary of the Process and Environmental Monitoring Methods Working Group Meeting with the FDA Emerging Technology Team

The Process and Environmental Monitoring Methods Working Group, composed of members from industry and instrument manufacturers, met with the FDA Emerging Technology Team to discuss bio-fluorescent particle counting technology, a type of rapid microbiological method. This is a summary of the meeting including submitted questions and answers, and the Process and Environmental Monitoring Methods Working Group's understanding of the FDA Emerging Technology Team's points made.

Rapid microbiological testing method

Classical microbiological methods currently have unacceptably long cycle times. Rapid microbiological strategies are accessible within the marketplace for about 10 years and are mostly used in the clinical laboratory and in food industries. However, their reapplication in the pharma industry has wide range of advantage. A comparison with ancient strategies to be conjointly performed. During this review, data concerning the validation of RMM strategies described within the document was given in addition as proof of the issue of validation of those strategies. A comparison with ancient strategies is additionally mentioned. This data is beneficial to the industries and in the labs which will doubtless be adopted. These strategies for microorganism free products. This methodology for microorganism identification will be delicate, accurate and fast. How the laboratory should be maintained for carrying out different tests, there should be good hygienic condition maintained. This article also includes different methods for identifying bacteria which is present in drug products as well as the material which are used for doing test. Presence of bacteria may affect the activity of drug product and bio availability may decreases and potency the drug may loss. How the laboratory should be maintained for carrying out different tests, there should be good hygienic condition maintained.

Case study on the environmental monitoring for biological manufacturing using Time-lapse Shadow Image Analysis

Bioburden control in the manufacturing facility is a serious concern regarding biologics due to the possibility of an significant economic impact due to batch failure from a bioburden incident. As a case study on effectively establishing a microbiological environmental monitoring program for cleanrooms, we focused on Time-lapse Shadow Image Analysis as a kind of Rapid Microbiological Method. In this study, the superior rapidity and accuracy were indicated for reference strains and environmental microbial on both 90 mm plate and RODAC plate at 25–30 °C. Especially superior performance in the counting was observed for B.subtilis, P.aeruginosa and A.brasiliensis. The first and the median of colony detection speed for environmental microbial were 12 h and 26 h, respectively. The colony detection rate was 90% at 40 h incubation. Additionally, the characterization of swarming behavior was recognized based on time-lapse image acquisition data at 30 min intervals. This case study indicated that the application for environmental monitoring can contribute to reducing the bioburden excursion risk due to both the rapid detection of colonies and real-time detection for swarming behavior. TSIA would be more acceptable and easier option for biologics due to providing simple interpretations for the results and reducing the time consumption.