Paenibacillus macerans can cause spoilage of milk during extended storage. However, the natural milk microbiota interferes with the enumeration of Paenibacillus species in raw milk. In this study, a qualitative SYBR Green real-time PCR assay based on the groEL gene was developed for detecting P. macerans (PMassay) in raw milk and compared with one designed for total Paenibacillus detection (TPassay). The specificity of the PMassay was confirmed against a panel of dairy-related spore forming isolates. In the presence of background DNA substituted up to 95%, P. macerans DNA could still be detected by the PMassay although interference occurred as non-target DNA substitution increased. The PMassay was sensitive (detection limit of 2 log CFU/ml in milk) and specific as non-P. macerans isolates gave a Ct > 30. After enrichment of raw milk for 7 days at 37 °C in Reinforced Clostridial Medium with D-cycloserine (RCM-D) under anaerobiosis, Paenibacillus was detected in 10 of the 16 raw milk samples tested. Enrichment in RCM-D yielded about 0.5 to 5.8 log CFU/ml total Paenibacillus and 0.3 to 4.6 log CFU/ml P. macerans in the samples. The assay could be useful in commercial settings, allowing a sensitive detection of P. macerans.

变形杆菌可能会在长期储存中导致牛奶变质。但是，天然牛奶菌群会干扰原奶中Paenibacillus物种的计数。在这项研究中，开发了一种基于groEL基因的定性SYBR Green实时PCR检测试剂盒，用于检测原料奶中的Macerans假单胞菌（PMassay），并将其与专为总Paenibacillus检测设计的一种（TPassay）进行比较。相对于一组与乳制品相关的孢子形成分离物，证实了PMassay的特异性。在存在背景DNA的情况下，最高可替代95％的鲭鱼尽管由于非靶DNA取代的增加而产生干扰，但PMassay仍可检测到DNA。所述PMassay是敏感（2个对数的检测极限CFU在牛奶/ ml）的具体为非和P.浸分离物在37℃下在强化梭菌培养基给予了的Ct> 30的原料奶7天富集后用D-在厌氧的环丝氨酸（RCM-D）中，在测试的16个原奶样品中的10个中检出了Paenibacillus。在样品中，RCM-D的富集产生约0.5至5.8 log CFU / ml的总杆状芽胞杆菌和0.3至4.6 log CFU / ml的南美白生菌。该测定法在商业环境中可能有用，可以灵敏地检测出鲭鱼。