The benefit of culture-independent methods to detect bacteria and fungi in re-infected root filled teeth: a pilot study.

No abstract

A patient with oral squamous cell carcinoma (OSCC) underwent complex surgical tumor therapy, including the reconstruction of soft tissues using a radial forearm flap. Due to venous congestion that could only partly be resolved by revision surgery, leech therapy was started on the second postoperative day. The patient developed pneumonia and sepsis and died as a result of septic shock, despite having received targeted broad-spectrum antibiotic therapy since day 5. Aeromonas spp. were cultured from both the patient’s specimens and unused leeches. Biochemical identification and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) yielded inconsistent identification results. Finally, microbiological identification of Aeromonas spp. was performed via 16S rDNA sequencing and use of the basic local alignment search tool (BLAST), and strains from both the patient and the leeches were identified as Aeromonas veronii. Aeromonas spp. strains derived from the patient and leeches and independent laboratory strains were submitted to randomly amplified polymorphic DNA (RAPD) subtyping. RAPD of A. veronii strains from both sources revealed an identical pattern, strongly suggesting the transmission of A. veronii from the leeches to the patient. Physicians should be aware of the potential for severe lethal infections as a fatal side-effect of leech therapy in critically ill patients, which should be addressed using antibiotic prophylaxis.

Purpose Acanthamoeba keratitis (AK) is a painful and possibly sight-threatening ocular infection. While the correct diagnosis and specific treatment in the early stages significantly improve the prognosis, the disease is often misdiagnosed and in clinical examination confused with other forms of keratitis. Polymerase chain reaction (PCR) for the detection of AK was first introduced in our institution in December 2013 to improve the timely diagnosis of AK. The aim of this study was to assess the impact of implementation of Acanthamoeba PCR on the diagnosis and treatment of the disease in a German tertiary referral center. Patients and methods Patients treated for Acanthamoeba keratitis between 1st of January 1993 and 31st of December 2021 in the Department of Ophthalmology of the University Hospital Duesseldorf were identified retrospectively via in-house registries. Evaluated parameters include age, sex, initial diagnosis, method of correct diagnosis, duration of symptoms until correct diagnosis, contact lens use, visual acuity, and clinical findings as well as medical and surgical therapy by keratoplasty (pKP). In order to assess the impact of implementation of Acanthamoeba PCR, the cases were divided into two groups (before (pre-PCR group) and after PCR implementation (PCR group). Results Seventy-five patients with Acanthamoeba keratitis were included (69.3% female, median age 37 years). Eighty-four percent (63/75) of all patients were contact lens wearers. Until PCR was available, 58 patients with Acanthamoeba keratitis were diagnosed either clinically (n = 28), by histology (n = 21), culture (n = 6), or confocal microscopy (n = 2) with a median duration until diagnosis of 68 (18; 109) days. After PCR implementation, in 17 patients, the diagnosis was established with PCR in 94% (n = 16) and median duration until diagnosis was significantly shorter with 15 (10; 30.5) days. A longer duration until correct diagnosis correlated with a worse initial visual acuity (p = 0.0019, r = 0.363). The number of pKP performed was significantly lower in the PCR group (5/17; 29.4%) than in the pre-PCR group (35/58; 60.3%) (p = 0.025). Conclusions The choice of diagnostic method and especially the application of PCR have a significant impact on the time to diagnosis and on the clinical findings at the time of confirmation of diagnosis and the need for penetrating keratoplasty. In contact lens–associated keratitis, the first crucial step is to take AK into consideration and perform a PCR test as timely confirmation of diagnosis of AK is imperative to prevent long-term ocular morbidity.

Coagulase-negative staphylococci (CoNS), members of the skin commensal microbiota, are increasingly associated with local or systemic infections due to a shift in patient populations in recent decades. Subsequently, more CoNS strains have been subjected to antibiotic susceptibility testing (AST), thus leading to the increased detection of teicoplanin resistance. However, data concerning teicoplanin resistance among CoNS strains remain limited, heterogeneous, and inconclusive. We collected 162 consecutive CoNS strains identified using Vitek-2 as teicoplanin-resistant and tested them with a range of AST methods. The results of standard and high inoculum broth microdilution (sBMD; hBMD), agar dilution (AD) after 24 h and 48 h incubation, standard and macrogradient diffusion strip (sGDT, MET), screening agar, and disc diffusion were compared to assess their robustness and to establish a diagnostic algorithm to detect teicoplanin resistance. sBMD was used as the reference method, and the lowest number of strains were teicoplanin-resistant using this method. sGDT and disc diffusion generated similar results to sBMD. Compared with sBMD, AD-24 h generated the lowest number of false teicoplanin-resistant strains, followed by hBMD, AD-48 h, and Vitek-2. sGDT, a fast, easy, affordable method in diagnostic settings, generated the highest rate of false teicoplanin-susceptible strains. Vitek-2 testing produced the highest number of teicoplanin-resistant strains. Only in two strains was the initial Vitek-2 teicoplanin resistance confirmed using five other AST methods. In conclusion, the different antibiotic susceptibility testing methods generated inconsistent, inconclusive, and discrepant results, thus making it difficult to establish a diagnostic algorithm for suspected teicoplanin resistance. Teicoplanin testing proved to be challenging and easily influenced by technical factors. This study aimed not only to raise awareness of teicoplanin resistance testing but also of the need for future studies focusing on the clinical efficacy of teicoplanin in relation to its susceptibility results.