Bacterial DNA in the diagnosis of spontaneous bacterial peritonitis

Abstract

SUMMARY BackgroundDespite inoculation into blood culture bottles, ascitic fluid culture is negative in 50% of cases of spontaneous bacterial peritonitis (SBP).

Identification of Bacterial Pathogens in Ascitic Fluids from Patients with Suspected Spontaneous Bacterial Peritonitis by Use of Broad-Range PCR (16S PCR) Coupled with High-Resolution Melt Analysis

“…Our 16S PCR was able to detect bacterial DNA in 100% (17/17) of culture-positive samples, and HRMA was able to correctly identify the bacterial species identified by culture in 70.6% (12/17) of positive samples in our study. The rate of positivity for bacterial DNA, 19.8% (21/106), detected from ascitic fluid samples in our study is comparable with the rates in other reported studies (3,4,8,9,11,12,(19)(20)(21)29). The overall sensitivity for detecting the presence of eubacterial DNA in an ascitic fluid sample and the specificity for the 16S PCR were high, at 100% (17/17) and 91.5% (85/89), respectively.…”

Application of Qualitative and Quantitative Real-Time PCR, Direct Sequencing, and Terminal Restriction Fragment Length Polymorphism Analysis for Detection and Identification of Polymicrobial 16S rRNA Genes in Ascites

“…Application of culture-independent PCR-based methods for the detection of bacterial DNA (bactDNA) in ascites has been proposed as a suitable tool to improve pathogen identification in patients at risk for SBP (6) and has indicated that bactDNA derives in most cases from a single pathogen (7)(8)(9). However, recent studies using 16S rRNA-based fingerprinting analyses have shown that ascites may also be polymicrobial (10,11) and that the bacterial spectrum is broader than that previously reported in context with SBP (11).…”

The Role of DNA Amplification and Cultural Growth in Complicated Acute Appendicitis

“…15,16 Similar results were available for microbiological diagnosis in spontaneous bacterial peritonitis where RT-PCR increased the efficacy of cultures: RT-PCR detected pathogen in nearly all the patients with positive culture, and RT-PCR resulted positive in half of the patients with negative cultures. 17 Although molecular diagnosis cannot be considered as a replacement method in the clinical setting, there is an increasing awareness of the benefits, which may derive from the combined use of molecular analyses and conventional microbiology to optimize the research of pathogens. 16,24,25 Concerning our pilot study, RT-PCR has been undoubtedly a valuable tool to detect aggressive pathogens such as anaerobia, which are usually difficult to preserve alive in biological cultures.…”

“…[15][16][17][18][19] In pleural empyema, RT-PCR played an important role identifying pathogens in those patients with no culture growth, who had followed a previous antibiotic course. 15,16 Similar results were available for microbiological diagnosis in spontaneous bacterial peritonitis where RT-PCR increased the efficacy of cultures: RT-PCR detected pathogen in nearly all the patients with positive culture, and RT-PCR resulted positive in half of the patients with negative cultures.…”

“…14 RT-PCR has been employed in several studies with pediatric populations confirming that a prompt detection of either viral or bacterial agent contributes to patient care, which in turn has effect on both hospital stay and antibiotic usage. [15][16][17][18][19] To date, a couple of studies about the use of RT-PCR to detect pathogens in the peritoneal fluid during acute appendicitis have been carried out without any further conclusion. 20,21 Taking into account these previous reports, we set a pilot study in order to better understand and compare the potential advantages of DNA amplification versus traditional bacterial growth culture in complicated acute appendicitis, and identify, when possible, those categories of patients, who might benefit from this combined approach.…”