Singh Ashutosh, Sharma Brijesh, Mahto Kaushal Kumar, Meis Jacques F, Chowdhary Anuradha
Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi 110 007, India.
Department of Medicine, Post Graduate Institute of Medical Education and Research, Dr. Ram Manohar Lohia Hospital, Delhi 110 001, India.
J Fungi (Basel). 2020 May 20;6(2):67. doi: 10.3390/jof6020067.
Aspergillosis due to azole-resistant is a worldwide problem with major therapeutic implications. In patients with invasive aspergillosis, a low yield of fungal cultures results in underestimation of azole resistance. To detect azole resistance in , we applied the AsperGenius Resistance multiplex real-time polymerase chain reaction (PCR) assay to detect TR/L98H, and TR/T289A/Y121F mutations and the AsperGenius G54/M220 RUO PCR assay to detect G54/M220 mutations directly in bronchoalveolar lavage (BAL) samples of 160 patients with chronic respiratory diseases in Delhi, India. Only 23% of samples were culture-positive compared to 83% positivity by species PCR highlighting concerns about the low yield of cultures. Notably, 25% of BAL samples (33/160 patients) had azole resistance-associated mutation by direct detection using PCR assay. Detection of resistance-associated mutations was found mainly in 59% and 43% patients with chronic pulmonary aspergillosis (CPA) and allergic bronchopulmonary aspergillosis (ABPA), respectively. Overall, a G54 mutation, conferring itraconazole resistance, was the predominant finding in 87.5% and 67% of patients with CPA and ABPA, respectively. In culture-negative, PCR-positive samples, we detected azole-resistant mutations in 34% of BAL samples. Azole resistance in chronic diseases remains undiagnosed, warranting standardization of respiratory culture and inclusion of rapid techniques to detect resistance markers directly in respiratory samples.
耐唑类曲霉病是一个具有重大治疗意义的全球性问题。在侵袭性曲霉病患者中，真菌培养阳性率低导致对唑类耐药性的低估。为了检测曲霉的唑类耐药性，我们应用AsperGenius耐药多重实时聚合酶链反应（PCR）检测法来检测TR/L98H和TR/T289A/Y121F突变，并应用AsperGenius G54/M220 RUO PCR检测法直接在印度德里160例慢性呼吸道疾病患者的支气管肺泡灌洗（BAL）样本中检测G54/M220突变。与曲霉菌种PCR 83%的阳性率相比，只有23%的样本培养呈阳性，这凸显了对培养阳性率低的担忧。值得注意的是，通过PCR检测法直接检测发现，25%的BAL样本（33/160例患者）存在与唑类耐药相关的突变。耐药相关突变的检测主要分别在59%的慢性肺曲霉病（CPA）患者和43%的变应性支气管肺曲霉病（ABPA）患者中发现。总体而言，赋予伊曲康唑耐药性的G54突变分别在87.5%的CPA患者和67%的ABPA患者中是主要发现。在培养阴性、PCR阳性的样本中，我们在34%的BAL样本中检测到了唑类耐药突变。慢性曲霉病中的唑类耐药性仍未得到诊断，需要规范呼吸道培养并纳入快速技术以直接在呼吸道样本中检测耐药标志物。
