Aran Gemma, Pleguezuelos Vanessa, Blanco Margarita, Garcia Cristina, Jallow Mariama, López Mar, Monge Sara, Casamitjana Natalia, Alonso-Nogués Eva, Soria Gloria
Banc de Sang i Teixits (Blood and Tissue Bank of Catalonia), 08005 Barcelona, Spain.
Grup de Medicina Transfusional, Vall d'Hebron Institut de Recerca (VHIR), Vall d'Hebron Hospital Universitari, 08035 Barcelona, Spain.
Microorganisms. 2025 Jul 11;13(7):1640. doi: 10.3390/microorganisms13071640.
Donor human milk (DHM) can harbor microbial contaminants that cause serious infections in premature infants. is a pathogen frequently found in DHM, capable of forming spores that can resist Holder pasteurization (62.5 °C, 30 min). Since no microbial growth is acceptable in post-pasteurized DHM, microbiological testing of pre-pasteurized DHM provides information about its contamination level to determine if it should be accepted for pasteurization. Culture is the gold standard in microbiological control but it requires 24-48 h to provide results. In this study we developed and validated a non-commercial real-time PCR assay for the detection of (BC test) in DHM specimens on a fully automated high-throughput platform, the cobas 6800 system. The BC test showed excellent sensitivity and specificity, repeatability and linearity over an 8-log range and a low limit of detection in milk specimens, as well as good agreement with selective culture methods. BC test was then used to systematically control all milk donations (3439) over a 24-month period. was detected in 14.2% of DHM, with monthly rates ranging from 6 to 29% and a significantly higher incidence in warmer months. Incorporating this assay into our laboratory workflow improved efficiency and reduced turnaround time.
供体人乳（DHM）可能含有会导致早产儿严重感染的微生物污染物。 是DHM中经常发现的一种病原体，能够形成可抵抗 Holder 巴氏杀菌法（62.5°C，30分钟）的孢子。由于巴氏杀菌后的DHM中不允许有微生物生长，因此对巴氏杀菌前的DHM进行微生物检测可提供其污染水平的信息，以确定是否应接受其进行巴氏杀菌。培养是微生物控制的金标准，但需要24 - 48小时才能得出结果。在本研究中，我们在全自动高通量平台 cobas 6800系统上开发并验证了一种用于检测DHM样本中 （BC检测）的非商业实时PCR检测方法。BC检测在8个对数范围内显示出优异的灵敏度、特异性、重复性和线性，在牛奶样本中的检测限较低，并且与选择性培养方法具有良好的一致性。然后，BC检测被用于系统地控制24个月内的所有母乳捐赠（3439份）。在14.2%的DHM中检测到了 ，月发生率在6%至29%之间，在温暖月份的发生率显著更高。将该检测方法纳入我们的实验室工作流程提高了效率并缩短了周转时间。
