Diagnostic Value of 16S Ribosomal RNA Gene Polymerase Chain Reaction/Sanger Sequencing in Clinical Practice. (18th August 2021)

• Record Type: Journal Article
• Authors: Fida, Madiha; Khalil, Sarwat; Abu Saleh, Omar; Challener, Douglas W; Sohail, Muhammad Rizwan; Yang, Joshua N; Pritt, Bobbi S; Schuetz, Audrey N; Patel, Robin
• Is Part Of: Clinical infectious diseases. Volume 73:Number 6(2021)
• Page Start: 961
• Page End: 968
• Publication Date: 2021-08-18
• Subjects: molecular diagnostics -- 16S rRNA gene PCR -- Sanger sequencing -- broad range bacterial PCR -- bacterial infections; Communicable diseases -- Periodicals; 616.905
• DOI: 10.1093/cid/ciab167
• Languages: English
• ISSNs: 1058-4838

Abstract

Abstract: Background: Accurate microbiologic diagnosis is important for appropriate management of infectious diseases. Sequencing-based molecular diagnostics are increasingly used for precision diagnosis of infections. However, their clinical utility is unclear. Methods: We conducted a retrospective analysis of specimens that underwent 16S ribosomal RNA (rRNA) gene polymerase chain reaction (PCR) followed by Sanger sequencing at our institution from April 2017 through March 2019. Results: A total of 566 specimens obtained from 460 patients were studied. Patients were considered clinically infected or noninfected based on final diagnosis and management. In 17% of patients, 16S rRNA PCR/sequencing was positive and in 5% of patients, this test led to an impact on clinical care. In comparison, bacterial cultures were positive in 21% of patients. Specimens with a positive Gram stain had 12 times greater odds of having a positive molecular result than those with a negative Gram stain (95% confidence interval for odds ratio, 5.2–31.4). Overall, PCR positivity was higher in cardiovascular specimens (37%) obtained from clinically infected patients, with bacterial cultures being more likely to be positive for musculoskeletal specimens ( P < .001). 16S rRNA PCR/sequencing identified a probable pathogen in 10% culture-negative specimens. Conclusion: 16S rRNA PCR/sequencing can play a role in the diagnostic evaluation of patients with culture-negative infections, especially those with cardiovascular infections. Abstract : Our study suggests the 16S ribosomal RNA gene polymerase chain reaction and Sanger sequencing is a clinically useful test and can play an important role in diagnostic evaluation of patients with culturenegative infections, especially with cardiovascular infections.