Pawar Megha Yogesh, Hatolkar Swarupa Mahesh, Misra Rabindra Nath
Lecturer (Microbiology), Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth (DPU), Tathawade, Pune, India.
Assistant Professor (Microbiology), Dr. D. Y. Patil Medical College, Hospital and Research Centre, Dr. D. Y. Patil Vidyapeeth (DPU), Pimpri, Pune, India.
Med J Armed Forces India. 2022 Jul;78(3):271-276. doi: 10.1016/j.mjafi.2020.03.023. Epub 2020 Jul 10.
Since the last two decades, substantial increase in fungal infections has been observed in immunocompromised hosts. Virulence factors of play a role in adherence, haemolytic activity, phenotypic switching and production of hydrolytic enzymes. The secreted aspartyl proteinases ( family) contribute to adhesion, tissue damage and invasion, while phospholipase () supports the hydrolysis of phospholipids. Very few studies showed the correlation of phenotypic activity and detection of genes contributing to the similar enzyme activity. Therefore, our study aimed at demonstrating correlation between phenotypic production of phospholipase and proteinase enzymes with genetic level detection of and genes.
The present study was carried out on a total of 799 samples over a period of one year. Culturing was carried out on Sabouraud's dextrose agar. Confirmation and speciation of spp. was carried out using cornmeal agar and CHROMagar and by the germ tube test, urease test and automated identification system Vitek-2, and antifungal susceptibility testing was performed on isolates. Phenotypic phospholipase and proteinase activity was analysed, and molecular detection of and genes was carried out.
In our study, we have screened 799 samples for mycological protocol; of which, 269 (33.6%) were species, 44% (119) of which were . Proteinase activity was exhibited by 77 (64.7%) and phospholipase activity was exhibited by 73 (61.34%) isolates, while 46.2% exhibited both activities among the species. The gene was detected in 97.3% isolates, while 4 was detected in 94.7% of isolates.
The study of in vitro expression of virulence factors and gene detection of will help to improve the prognosis despite the nature of the sample.
在过去二十年中，免疫功能低下宿主中的真菌感染显著增加。其毒力因子在黏附、溶血活性、表型转换和水解酶产生中起作用。分泌型天冬氨酸蛋白酶（家族）有助于黏附、组织损伤和侵袭，而磷脂酶（）支持磷脂的水解。很少有研究表明表型活性与导致相似酶活性的基因检测之间的相关性。因此，我们的研究旨在证明磷脂酶和蛋白酶的表型产生与和基因的基因水平检测之间的相关性。
本研究在一年时间内共对799份样本进行。在沙氏葡萄糖琼脂上进行培养。使用玉米粉琼脂、显色培养基以及芽管试验、尿素酶试验和自动鉴定系统Vitek-2对菌种进行确认和鉴定，并对分离株进行抗真菌药敏试验。分析磷脂酶和蛋白酶的表型活性，并进行和基因的分子检测。
在我们的研究中，我们按照真菌学方案筛选了799份样本；其中，269份（33.6%）为菌种，其中44%（119份）为。77份（64.7%）分离株表现出蛋白酶活性，73份（61.34%）分离株表现出磷脂酶活性，而在菌种中，46.2%的分离株同时表现出这两种活性。97.3%的分离株检测到基因，而94.7%的分离株检测到4基因。
尽管样本性质不同，但对毒力因子的体外表达和基因检测的研究将有助于改善预后。
