Application of Rapid Microbiological Assay in Tumor-Infiltrating Lymphocyte(TIL)Products
Objective:To evaluate sterility detection of tumor-infiltrating lymphocyte(TIL)products by means of respiratory signal-ing method and nucleic acid amplification(QPCR)technology method,to develop an efficient and reliable method for rapid microbial detection of immune cell products.Methods:Live immune cells with short shelf life and limit cell valume,cannot be sampled with tradi-tional microbial detection methods,"This study evaluated sterility detection of tumor-infiltrating lymphocyte(TIL)products by means of respiratory signaling and nucleic acid amplification technology,aiming to develop an efficient and reliable method for rapid microbial detection of immune cell products.With the verification of the 6 strains specified in the 1101 section of 2020 edition of Chinese Pharmaco-poeia,we extract immune cells from the patient's peripheral blood or tumor tissue,andtest TIL product in direct inoculation method of Chinese Pharmacopoeia,respiratory signal method and QPCR method seperately.After in vitro activation,the tumor cells were infused back into the body to kill the tumor cells,at the same time,the immune system of the patients was further activated to achieve the goal of survival or cure with the tumor.Results:Under the condition of<100 CFU·mL-1,the respiratory test result showed that the positive sig-nal was within 0.40～1.53 d.While the positive signal was detected within 4 h by QPCR method.There are no components interfering with the detection of bacteria and fungi in TIL products.The two methods have good specificity and durability with the respiratory signal method having the best detection limit and the QPCR method having the shortest detection time.Conclusion:The sensitivity and culture time of respiratory signal method are superior to the traditional direct inoculation method of Chinese Pharmacopoeia.QPCR reduces the detection time from 14 d to 4 hs.Both methods have good specificity.Therefore,they can be used as a rapid detection method under dif-ferent product conditions such as fresh or frozen,and has good application prospects in microbial testing of cell therapy products.
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