Sandeep kumar’s Post

Why Do We Incubate BET Reaction Tubes at 37°C for 60 Minutes? In the Bacterial Endotoxins Test (BET), incubating reaction tubes at 37°C ± 1 for exactly 60 ± 2 minutes is a key step. But why such specific conditions? Let's dive in. 1. Optimal Enzyme Activity The heart of BET lies in the Limulus Amebocyte Lysate (LAL) reagent, sourced from horseshoe crab blood. This enzyme reacts with bacterial endotoxins (LPS) at body temperature, 37°C, its peak activity zone. This ensures that the reaction happens consistently and at a rate that's both efficient and predictable. 2. Simulating Physiological Conditions By setting the incubation to body temperature, we're not just promoting enzyme activity; we're mirroring human physiological conditions. This alignment provides more accurate insights into how endotoxins might behave in a real-life scenario, such as within the human body. 3. Standardization for Reproducibility Consistency is crucial. 60 ± 2 minute gives a standardized period for reaction development. This precision means any detectable changes (color shifts, gel formation) are truly due to endotoxins, not incubation variables. 4. Complete Reaction Development For full sensitivity, sufficient time is essential. The 60-minute window ensures full reaction progression, allowing for accurate endotoxin detection. Shorter times could cause false negatives, while longer could risk overdevelopment. 5. Minimizing Variability A stable 37°C ± 1 incubation minimizes fluctuations, helping maintain reagent stability and preserving test integrity across runs. This carefully calibrated process ensures that BET is reliable and reproducible. As per ISO 23500/IS 17646 standards, particularly in applications like dialysis water testing, the endotoxin level must stay below 0.25 EU/ml to meet stringent safety requirements. The USFDA, too, recognizes BET's value in its sensitivity and reproducibility. The United States Pharmacopeia (USP) sets endotoxin limits for water types: 0.25 EU/ml for Water for Injection. The Indian Pharmacopeia also endorses BET for pyrogen-free assurance. ## microbiologist Sandeep Mishra ## #bet #microbiology #biology #iso #usfda# EU, GMP #

Clinical Microbiology is a Rapidly Changing and Upgrading Field and One Must Stay Abreast with Developments in the Areas of their Interest. Glad to Share the Certificate of Completion for CLSI M100 S34 Changes in 2024. #Antimicrobials #Antibiotics #antimicrobialstewardship

In microbiology, CFU stands for Colony Forming Units, a crucial measurement used to estimate the number of viable bacteria or fungal cells in a sample. This unit helps scientists quantify bacterial growth, assess contamination levels, and determine the efficacy of antimicrobial treatments. Each CFU represents a single viable organism capable of dividing and forming a colony, allowing researchers to calculate bacterial density in a sample. CFU counts are widely used in fields like environmental testing, food safety, pharmaceuticals, and clinical diagnostics. Measuring CFUs provides insight into the health and behavior of microbial populations, guiding decisions in healthcare, manufacturing, and research. #Microbiology #CFU #BacterialGrowth #Science #Healthcare #Research

𝗠𝗶𝗰𝗿𝗼𝗯𝗶𝗼𝗹𝗼𝗴𝘆 𝟭𝟬𝟬 𝗶𝗻𝘁𝗲𝗿𝘃𝗶𝗲𝘄 𝗾𝘂𝗲𝘀𝘁𝗶𝗼𝗻𝘀 𝟵𝟵. 𝗪𝗵𝘆 𝗱𝗼𝗲𝘀 𝘁𝗵𝗲 𝗺𝗲𝗱𝗶𝗮 𝗰𝗼𝗼𝗹 𝘁𝗼 𝟰𝟱 𝗱𝗲𝗴𝗿𝗲𝗲𝘀 𝗯𝗲𝗳𝗼𝗿𝗲 𝗽𝗼𝘂𝗿𝗶𝗻𝗴 𝘁𝗵𝗲 𝗽𝗹𝗮𝘁𝗲𝘀? In microbiology, media is allowed to cool to around 45°C before pouring plates to prevent condensation and maintain the viability of any added supplements. If the media is too hot when poured, it can cause excessive condensation on the Petri dish lid as it cools. This moisture can drip onto the agar surface, potentially spreading bacteria or other organisms across the plate and disrupting isolated colony formation. Some media require the addition of heat-sensitive supplements (e.g., antibiotics) to support the growth of specific organisms. If the media is too hot, these supplements may degrade, reducing their effectiveness and potentially affecting the growth conditions. At 45°C, the agar cools at a rate that allows for uniform solidification, creating a flat, smooth surface ideal for streaking or spreading microorganisms. #microbiology #pharma #media #vijaykumarh

Last month was in microbiology department: Devices I used : Bact alert and BD bactec for blood culture container came Aerobic and anaerobic Cepheid use for PCR in MRSA and TB PCR Micro scan waik away use for reading ,chemical and antibiotic must prepare sample and use to panels (+) and (-) If there is growth in blood agar only use panel (+) If there is growth in blood and mac use panel (-) Reading: Gram(+) use catalase ,coagulase test S.aureus (+),(+) and colony appears Use antibiotic Opt in chocolate agar for S.pneumonia for sample CSF,sputum, Gram (-) Lactose fermentation in Mac will appear pink E.coli will be pink and small Klebsiella will appear pink and large mucoid

𝗪𝗵𝘆 𝘆𝗼𝘂 𝘂𝘀𝗶𝗻𝗴 𝘀𝗮𝗹𝗶𝗻𝗲 𝗳𝗼𝗿 𝘀𝘄𝗮𝗯𝘀? Saline is commonly used for swabs in microbiology and clinical applications because it provides an isotonic environment that preserves cell integrity. 𝗠𝗮𝗶𝗻𝘁𝗮𝗶𝗻𝘀 𝗠𝗼𝗶𝘀𝘁𝘂𝗿𝗲 Saline keeps the swab moist, preventing the sample from drying out, which helps in recovering microorganisms or cells during testing. 𝗣𝗿𝗲𝘀𝗲𝗿𝘃𝗲𝘀 𝗩𝗶𝗮𝗯𝗶𝗹𝗶𝘁𝘆 It doesn’t kill or alter the microorganisms, ensuring an accurate sample for culturing or analysis. 𝗖𝗼𝗺𝗽𝗮𝘁𝗶𝗯𝗶𝗹𝗶𝘁𝘆 Being isotonic (same salt concentration as cells or tissues), saline prevents osmotic stress that could damage the microorganisms or cells being collected.

5 Characteristics Of Forward-Thinking Microbiology Labs In 2025 https://lnkd.in/eVnR9fdd - By Tim Sandle, Ph.D., CBiol, FIScT. This article assesses five trends that are likely to be part of the cost and efficiency focus during 2025.

𝗪𝗵𝘆 𝘆𝗼𝘂 𝘂𝘀𝗶𝗻𝗴 𝘀𝗮𝗹𝗶𝗻𝗲 𝗳𝗼𝗿 𝘀𝘄𝗮𝗯𝘀? Saline is commonly used for swabs in microbiology and clinical applications because it provides an isotonic environment that preserves cell integrity. 𝗠𝗮𝗶𝗻𝘁𝗮𝗶𝗻𝘀 𝗠𝗼𝗶𝘀𝘁𝘂𝗿𝗲 Saline keeps the swab moist, preventing the sample from drying out, which helps in recovering microorganisms or cells during testing. 𝗣𝗿𝗲𝘀𝗲𝗿𝘃𝗲𝘀 𝗩𝗶𝗮𝗯𝗶𝗹𝗶𝘁𝘆 It doesn’t kill or alter the microorganisms, ensuring an accurate sample for culturing or analysis. 𝗖𝗼𝗺𝗽𝗮𝘁𝗶𝗯𝗶𝗹𝗶𝘁𝘆 Being isotonic (same salt concentration as cells or tissues), saline prevents osmotic stress that could damage the microorganisms or cells being collected.

Systemic Microbiology Simplified!

Food for thought. In the international market, there are roughly three widely recognized brands of manufacturers for automated susceptibility testing systems, as well as numerous Chinese solutions from various manufacturers. All of them operate on a standard protocol, delivering results within 18-24 hours. Since 2018, there has been a trend towards accelerated susceptibility testing methods directly from blood cultures or pure cultures within 2-4 hours. Currently, there are about nine ready-to-use solutions on the market, not counting Chinese ones (I know three or four of them). This suggests that in the next two years, both the market and the microbiology service itself could change significantly..... #microbiology #RAST #AST