ICU care through NGS – Based identification of infectious agents: A comparative study

• Published in: Heliyon 2024-07, Vol.10 (14), p.e34538, Article e34538
• Main Authors: Ramanathan, Aishwariya, Marimuthu, Anusuya, Abayasekara, Lalanika M., Jeyaratnam, Thurga, Chandrasekharan, Vishvanath, Perera, Jennifer, Ramasubramanian, Venkatasubramanian, Sethuraman, Nandini, Samarasinghe, Sumudu, Peiris, Varuna, Gnanam, Vaz S.
• Format: Article
• Language: English
• Subjects: 16s rRNA sequencing; comparative study; diagnostic techniques; fungal culture; genomics; India; infectious diseases; ITS sequencing; Metagenomics; microbiology; mortality; Next generation sequencing; Pan bacterial identification; Pan fungal identification; pathogens; pollution; Sepsis; statistics
• ISSN: 2405-8440; 2405-8440
• DOI: 10.1016/j.heliyon.2024.e34538

Sepsis claims 1 in 5 lives annually as per global statistics. Sepsis incidence in recent studies represents at least 35 % of all ICU admissions and has a high mortality rate, especially in the presence of co-existing morbidities. The challenge has been to accurately diagnose the causative organism, considering factors such as possible polymicrobial infections, commensals and environmental contaminants. Legacy techniques such as culture, automated culture systems or even newer species-specific PCR or film array these challenges difficult to overcome. The Bactfast® and Fungifast® assays along with the integrated workflow is based on next generation sequencing and have the ability to demarcate infecting pathogen from contamination and commensal. The unique ability to pinpoint the exact pathogen, considering the commensal and contamination in a variety of samples, with an extremely high sensitivity could lead it to be a tool of diagnostic choice for non-resolving ICU sepsis due to its comprehensive coverage and speed. The aim of this study was to evaluate the use of Bactfast® and Fungifast® as a last mile diagnostic tool in a ICU setting. This study was carried out considering access to four intensive care units (ICU). Legacy testing, mostly done on culture, was conducted at the various integrated microbiology facilities of the hospitals where the ICUs were located, in Chennai, India. NABL accredited laboratory Micro Genomics (India) Pvt Ltd, was established as the central processing facility for next generation sequencing to run the Bactfast® and Fungifast® assay. Co-relation of results for 490 samples was done retrospectively by a multi-disciplinary team of consultants which comprised of microbiologists, and infectious disease physicians. The diagnostic workflow established with the Bactfast® assay provided a sensitivity of 94.1 % and specificity of 86.6 %. Identification of pathogens in Bactfast® was better when compared to the data published in 2017, as reflected by positive co-relation with clinical confirmation. Although the Fungifast® specificity was high, at 99.4 %, only 12 samples were positive on fungal culture out of 490 samples. Therefore, it was concluded a further study for fungi based on multiple technologies with more true positive samples is required to evaluate the test. Bactfast® can identify pathogens in a sample without any bias. Its introduction as diagnostic modality in life threatening ICU sepsis could reduce mortality and morbidity. Al