Application of Rapid Biological Indicators Coupled With Auto-Reader for the Quality Assurance of Surgical Instruments After Sterilization at a Cardiac Hospital in Bangladesh.

Background Sterilization failure is one of the main reasons for surgical site infections (SSIs). The biological indicator (BI) test is the most reliable test to check sterilization efficiencies. But 48 hours BI test result makes the monitoring process time-consuming. Rapid BI testing can be time demanding in this regard. Therefore, the objective is to determine the importance of rapid BI monitoring for the quality assurance of sterile surgical instruments. Methods This study was conducted in the Labaid Cardiac Hospital, Bangladesh from April 1, 2021, to July 8, 2021. A total of 100 steam and 100 ethylene oxide (EO) rapid BIs and an auto reader incubator were used to conduct this research. Quick BI of steam and EO were used once per day and tested by the auto reader. Later, all the tested BIs were incubated for 48 hours by a conventional incubator to confirm the auto reader's rapid BI test results. Result All the EO BI results were found negative, but the BI was found positive twice in steam sterilization. Surgical items of those two loads were re-sterilized. Again, after checking the BI result, the items were released. All BIs except positive steam rapid BIs were found with no growth after 48 hours of incubation for cross-checking of auto reader results. In positive rapid BI of steam, growth was found after 48 hours of incubation. Conclusion When sterilization failure occurred, process recall could not be possible at that time if rapid BI tests were not performed. So, integration of a rapid BI test with an auto reader can save the patient from critical SSI.

Studies of the impact of occupational exposure of pharmaceutical workers on the development of antimicrobial drug resistance.

OBJECTIVES: Pharmaceutical workers involved with the production of antimicrobial drugs are exposed to various antimicrobial chemicals in different steps of manufacturing such as grinding, sieving, compression, granulation, mixing and filling. These exposures may lead to the development of multidrug resistance (MDR) in bacteria. Scientific reports on the occupational health hazard of pharmaceutical workers involved in manufacturing antibiotics are scarce. The present study aimed to compare the degree of bacterial resistance in pharmaceutical workers in Bangladesh to that of individuals not involved in the pharmaceutical field. METHODS: Twenty male workers from five local pharmaceutical companies and twenty male subjects not involved in the pharmaceutical field (non-pharmaceutical subjects) were randomly selected. Nasal fluid, mucus/cough and stool specimens were collected from each subject and were cultured separately at 37°C for 24 hours to obtain bacterial growth. The cultured species were then identified, isolated and subjected to microbial sensitivity testing against 18 different antibiotics from eight different groups by the disk diffusion method. Staphylococcus spp., Pseudomonas spp. and Escherichia coli were identified and isolated from the culture of nasal fluids, mucuses and stools, respectively. RESULTS: All the isolated species of bacteria exhibited significant enhancement of the degree of MDR in pharmaceutical workers compared with non-pharmaceutical subjects. Workers with a longer working history had greater degree of antibiotic resistance and vice versa. It can be certainly considered that the exposure of pharmaceutical workers to antibiotic agents resulted in a high incidence of multidrug resistance. CONCLUSIONS: Effective steps should be taken to minimize inherent exposure of pharmaceutical workers to antibiotics during work to prevent antimicrobial drug resistance.

Direct construction of an open-sandwich enzyme immunoassay for one-step noncompetitive detection of thyroid hormone T4.

To establish a sensitive noncompetitive immunoassay for thyroxine (T4), we attempted to isolate anti-T4 antibodies from a phage display library based on a phagemid pDong1 ( Dong et al. Anal. Biochem.2009, 36, 386 ), which was designed to enable open-sandwich enzyme-linked immunosorbent assay (OS-ELISA) after selection on immobilized antigen. After the Fab-displaying phage library made from the splenocytes of T4-KLH immunized mice was subjected to biopanning on T4-BSA, two T4-specific clones were obtained. When they were assayed by indirect competitive ELISA, both clones showed low IC(50) (5-13 ng/mL), indicating their high affinity to T4. When they were used for OS-ELISA that detects antigen-dependency of the interaction between variable domains V(H) and V(L), a clone successfully detected 1 ng/mL of T4 with a working range superior to that of competitive IA. OS-ELISA was also performed with maltose binding protein (MBP)-fused V(H)/V(L) of this clone, which showed a detection limit less than 0.1 ng/mL T4. Moreover, the assay showed cross-reactivity with T3 similar to that of competitive ELISA, and also gave a reasonable total serum T4 concentration (90 ng/mL) from ethanol-extracted sample serum using the recombinant proteins. This is the first direct construction of an OS-ELISA system bypassing hybridoma, which will be applicable to the detection of many other small molecule antigens.

Micro open-sandwich ELISA to rapidly evaluate thyroid hormone concentration from serum samples.

BACKGROUND: Thyroxine (T4) is the most commonly measured thyroid hormone for the diagnosis of thyroid function. To elucidate a rapid and sensitive assay for T4, we made a microfluidics-based noncompetitive immunodetection chip system using anti-T4 antibody fragments obtained from a phage display library. RESULTS: Based on the open-sandwich ELISA principle that detects antigen-dependency of the interaction between the two antibody variable regions V(H) and V(L), we could detect less than 1 ng/ml of T4. The assay was also successfully applied to evaluate total T4 concentration in the serum of healthy individuals. CONCLUSION: This would be the first micro open-sandwich ELISA constructed with antibody fragments directly selected from immunized mice. The system will be applied to the sensitive detection of many diagnostic markers.