Phage-Mediated Molecular Detection (PMMD): A Novel Rapid Method for Phage-Specific Bacterial Detection

Viruses

Biswajit Biswas¹  Francisco Malagon¹  Luis A. Estrella¹  Theron Hamilton¹  Michael G. Stockelman²  Nimpha Teneza-Mora² 

[1] Genomics & Bioinformatics Department, Biological Defense Research Directorate, Naval Medical Research Center-Frederick, Fort Detrick, MD 21702, USA;Wound Infections Department, Infectious Diseases Directorate, Naval Medical Research Center-Silver Spring, Silver Spring, MD 20910, USA;

关键词: Bacteriophage;    phage;    RNA;    bacterial detection;    antibiotic resistance;    S. aureus;   

DOI  :  10.3390/v12040435

来源: DOAJ

【 摘 要 】

Bacterial infections pose a challenge to human health and burden the health care system, especially with the spread of antibiotic-resistant populations. To provide effective treatment and improved prognosis, effective diagnostic methods are of great importance. Here we present phage-mediated molecular detection (PMMD) as a novel molecular method for the detection and assessment of bacterial antibiotic resistance. This technique consists of a brief incubation, of approximately ten minutes, of the biological sample with a natural bacteriophage (phage) targeting the bacteria of interest. This is followed by total RNA extraction and RT-PCR. We applied this approach to Staphylococcus aureus (SA), a major causative agent of human bacterial infections. PMMD demonstrated a high sensitivity, rapid implementation, and specificity dependent on the phage host range. Moreover, due to the dependence of the signal on the physiological state of the bacteria, PMMD can discriminate methicillin-sensitive from methicillin-resistant SA (MSSA vs. MRSA). Finally, we extended this method to the detection and antibiotic sensitivity determination of other bacteria by proving PMMD efficacy for Bacillus anthracis.

【 授权许可】

Unknown