The Application Analysis of Multiplex Real-Time Polymerase Chain Reaction Assays for Detection of Pathogenic Bacterium in Peritoneal Dialysis-Associated Peritonitis

Abstract

Background/Aims: To estimate the clinical value of bacterial detection in peritoneal dialysis-associated peritonitis (PDAP) by multiplex real-time polymerase chain reaction (RT-PCR). This study was undertaken to evaluate multiplex RT-PCR for identifying clinically significant bacteria in PDAP. Methods: Seventy peritoneal dialysate specimens were collected and traditional bacterial culture and universal primer RT-PCR detection of the bacterial were used. Results: The positive rate of traditional culture method was 65.71% (46/70) and that of universal primer RT-PCR was 81.42% (57/70). For 6 clinical commonly pathogenic bacteria, multiplex, and monoplex RT-PCR all detected 38 positive ones within the 57 specimens that were detected positive by universal primer RT-PCR. The results of the 2 methods were completely identical. Detecting bacteria by universal primer PCR and Monoplex RT-PCR needs 4–5 and 6–9 h, respectively, while multiplex RT-PCR needs less than 3 h. Conclusion: Our results demonstrated that the multiplex RT-PCR can detect several kinds of bacteria simultaneously and it is also more practical and convenient than monoplex RT-PCR.

Pathogenic spectrum and risk factors of peritoneal dialysis-associated peritonitis: a single-center retrospective study

“…This phenomenon could be explained by repeated infection or previous use of antibiotics before culture [ 30 ]. Furthermore, G + were the dominant pathogenic bacteria, which is in agreement with previous findings [ 31 , 32 ]. The G + bacteria were represented by S. epidermidis , a coagulase-negative Staphylococcus that habitats on the skin.…”

The application of metagenomic next-generation sequencing for detection of pathogens from dialysis effluent in peritoneal dialysis–associated peritonitis

“…1 However, studies show that many centres fail to meet this standard. 2 –4 Recently, metagenomic next-generation sequencing (mNGS) has emerged as a powerful technique for detecting pathogens such as bacteria, viruses, fungi and parasites owing to its high-throughput capacity and fast turnaround time. Thus, use of mNGS has begun to improve the etiological diagnosis of infectious diseases, including sepsis, 5 pneumonia 6 and nervous system infections.…”

Αξιολόγηση της κλινικής σημασίας της ανίχνευσης βακτηριακού DNA στο αίμα και στο ασκιτικό υγρό με τη χρήση πραγματικού χρόνου αλυσιδωτής αντίδρασης πολυμεράσης (Real time polymerase chain reaction -PCR) σε νοσηλευόμενους κιρρωτικούς ασθενείς με ισχυρή υποψία λοίμωξης

“…Αλλαγές στη μεθοδολογία όπως μεταβολή της θερμοκρασίας φύλαξης ή της τεχνικής εξαγωγής του DNA μπορούν να περιορίσουν, σε άλλοτε άλλο βαθμό, αλλά πιθανώς όχι να εξαλείψουν το φαινόμενο [244,245]. Η χρήση πιο ευαίσθητων τεχνικών PCR,…”