Conventional and new technologies are available for the enumeration of Staphylococcus aureus. Here are some commonly used methods: Conventional Methods: 1. Agar Plate Counting: This is a traditional method where a sample is serially diluted and plated on agar plates, typically Mannitol Salt Agar (MSA), which selectively allows the growth of Staphylococcus aureus. After incubation, colonies are counted, and the viable cell count is determined. 2. Most Probable Number (MPN) Method: The MPN method is a statistical technique used for estimating the number of bacteria in a sample. It involves inoculating multiple tubes with different dilutions of the sample and assessing growth based on turbidity. The MPN is determined by comparing the results with a statistical table. New Technologies: 1. Polymerase Chain Reaction (PCR): PCR is a molecular technique used to amplify specific DNA sequences. In the case of Staphylococcus aureus enumeration, quantitative PCR (qPCR) can be used to target specific genes unique to Staphylococcus aureus. The number of bacterial DNA copies is then quantified, providing an estimation of the bacterial load. 2. Real-time PCR (RT-PCR): RT-PCR combines the amplification of PCR with real-time monitoring of the amplification process. It allows for the quantification of DNA or RNA in real-time, providing rapid and accurate results. RT-PCR can be used to detect and quantify Staphylococcus aureus in a sample. 3. Flow Cytometry: Flow cytometry is a technique that analyzes individual cells or particles as they pass through a laser beam. It can be used for the enumeration of Staphylococcus aureus by labeling the bacteria with fluorescent dyes or antibodies specific to the organism. The instrument then detects and counts the labeled bacteria, providing quantitative information. 4. Digital PCR (dPCR): dPCR is a method that partitions the sample into thousands of individual reactions and performs PCR amplification in each partition. The presence or absence of the target DNA is detected in each reaction, and the number of positive partitions is used to estimate the absolute quantity of the target DNA, giving an enumeration of Staphylococcus aureus cells. These new technologies offer advantages such as increased sensitivity, faster results, and the potential for automation. However, it's important to note that the choice of method depends on factors such as the purpose of enumeration, available resources, and the level of accuracy and precision required.