Real-time PCR versus MALDI-TOF MS and culture-based techniques for diagnosis of bloodstream and pyogenic infections in humans and animals.

AIMS: This study was applied to evaluate the usefulness of a high-throughput sample preparation protocol prior to the application of quantitative real-time PCR (qPCR) for the early diagnosis of bloodstream and pyogenic infections in humans and animals compared to matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and classical culture. METHODS AND RESULTS: Saponin-mediated selective host cell lysis combined with DNase-1 was applied for processing of whole blood and pus clinical samples collected from suspected cases of septicaemia and pyogenic infections in humans and animals. The pre-PCR processing strategy enabled the recovery of microbial cells with no changes in their colony forming units immediately after the addition of saponin. DNase-1 was efficient for removing the DNAs from the host cells as well as dead cells with damaged cell membranes. The metagenomic qPCR and MALDI-TOF MS could identify the bacterial community of sepsis at species level with a concordance of 97·37% unlike the conventional culture. According to qPCR results, Staphylococcus aureus (24·24%) was predominated in animal pyogenic infections, whereas Klebsiella pneumonia (31·81%) was commonly detected in neonatal sepsis. CONCLUSIONS: Saponin combined with DNase-1 allowed the efficient recovery of microbial DNA from blood and pus samples in sepsis using qPCR assay. SIGNIFICANCE AND IMPACT OF THE STUDY: Metagenomic qPCR could identify a broad range of bacteria directly from blood and pus with more sensitivity, higher discriminatory power and shorter turnaround time than those using MALDI-TOF MS and conventional culture. This might allow a timely administration of a prompt treatment.

Primary Ciliary Body Marginal Zone Lymphoma Presenting as Hemorrhagic Hypopyon.

PURPOSE: Our purpose is to report a patient with primary unilateral ciliary body marginal zone lymphoma who initially presented with hemorrhagic hypopyon. METHODS: Retrospective review of the clinical, imaging, and immunohistopathological features of the case was performed. RESULTS: A 59-year-old man was referred with right anterior uveitis of unknown etiology which was unresponsive to systemic treatment. Slit-lamp biomicroscopy showed normotensive hemorrhagic hypopyon in that eye. Anterior segment ultrasound biomicroscopy revealed an iridociliary mass lesion. Because an anterior chamber paracentesis was noncontributory, a diagnostic cyclectomy was performed. Histopathological evaluation showed that the neoplastic cells were positive for CD20, lambda light chain, and BCL 2. BCL 6, CD10, CD5, SOX11, kappa, and Cyclin D1 stains were negative. The final diagnosis was extranodal marginal zone lymphoma of the ciliary body. CONCLUSIONS: Although rare, ciliary lymphoma may be a cause of intractable anterior uveitis. Repeat biopsies could be carried out when there is a high level of clinical suspicion.

Multidisciplinary Diagnostic Approach in Intraocular Lymphoma Featuring Pseudo-hypopyon: Case Series and Literature Review.

PURPOSE: Diagnosis of intraocular lymphoma (IOL) is usually achieved by histopathological analysis. However, it may lead to inconclusive results due to the scarcity of malignant cells obtained by biopsy, hence leading to delayed diagnosis. We report two cases of IOL with pseudo-hypopyon, a rare feature of IOL, as their initial ocular feature, diagnosed using a multidisciplinary diagnostic approach. Common clinical features of IOL with pseudo-hypopyon were also investigated. METHODS: Retrospective case series and literature review. RESULTS: Two cases of IOL, a 78-year-old female and a 59-year-old male, whom had been diagnosed with systemic B-cell lymphoma developed pseudo-hypopyon and visual impairment during the course of their chemotherapy. Diagnosis of IOL was achieved from anterior chamber aspiration samples with supplementary diagnostic tools including flow cytometric immunophenotyping, interleukin and IgH gene rearrangement analysis in addition to the conventional histopathological analysis. Generally, pseudo-hypopyon was more commonly seen in secondary IOL and may associate with hyphema and high intraocular pressure. CONCLUSION: Pseudo-hypopyon is a rare feature of IOL, more commonly seen in secondary IOL, which can be accompanied by hyphema and high intraocular pressure. Supplementary diagnostic tools such as flow cytometric immunophenotyping, interleukin analysis, and immunogloblin H gene rearrangement analysis are useful for supporting the diagnosis of IOL with pseudo-hypopyon.

Case for diagnosis. Suppurative nodule or something deeper?

The authors report the case of a 62-year-old man with a history of total left hip arthroplasty nine years previously presenting with a large infiltrated plaque on the posterior area of the left thigh with three months of evolution without systemic symptoms or elevated inflammatory markers. Computed tomography of the left lower limb revealed a 12-centimeter linear extension of the lesion to the posterior part of the left proximal femur. Prosthesis joint infection, although rare, is a surgical complication to be taken into account, even if the surgery was performed many years before.

[The additional criterion for the diagnosis of purulent-destructive forms of acute pyelonephritis].

AIM: to develop an additional criterion for objective diagnosis of purulent-destructive forms of acute pyelonephritis. MATERIALS AND METHODS: A total of 35 blood samples from patients with acute pyelonephritis aged from 19 to 85 years (mean age was 52 years) were studied. For the analysis, a classical bacteriological method of isolating blood culture and a modern molecular genetic method of fluorescent in situ hybridization (FISH) using DNA probes complementary to species-specific regions of the 16S rRNA gene of microorganisms were used. The obtained results were processed using the Statistica 6.0 program. RESULTS: The bacteriological study showed that only in one patient (2.9%) a blood culture was positive, whereas with FISH, polymicrobial infection consisting of two associates was observed in five patients (14.3%), and 207 specific luminescence from different DNA probes were detected in blood samples. The associations of members of the Enterobacteriaceae with S. aureus were 2.4 times more frequent than associations of the Enterobacteriaceae with S. epidermidis, indicating a development of the polymicrobial infection and, accordingly, the development of purulent stage of acute pyelonephritis. CONCLUSION: For the diagnosis of acute pyelonephritis, we suggest to use an additional criterion, which consists in applying of FISH method for the detection and simultaneous identification of bacteria that were adhered on the surface of erythrocytes and located intracellularly. This method may allow for the differential diagnosis of serous and purulent stages of acute pyelonephritis.

Supposed endogenous endophthalmitis caused by Serratia marcescens in a cat.

An 8-year-old male neutered domestic shorthair cat was presented for evaluation of acute respiratory distress. Respiratory auscultation revealed a diffuse and symmetric increase in bronchovesicular sounds. Thoracic radiographs showed a diffuse unstructured interstitial pulmonary pattern with multifocal alveolar foci. Despite an aggressive treatment with supportive care, including oxygenotherapy and systemic antibiotics, progressive respiratory distress increased. Three days after the presentation, acute anterior uveitis was noticed on left eye. Ophthalmic examination and ocular ultrasonography revealed unilateral panuveitis with ocular hypertension. The right eye examination was unremarkable. Cytological examination of aqueous humor revealed a suppurative inflammation. Serratia marcescens was identified from aqueous humor culture. Primary pulmonary infection was suspected but was not confirmed as owners declined bronchoalveolar lavage. Active uveitis resolved and cat's pulmonary status improved after appropriate systemic antibacterial therapy. Vision loss was permanent due to secondary mature cataract. To the best of authors' knowledge, this is the first report of endogenous bacterial endophthalmitis secondary to S. marcescens infection in a cat.

Case for diagnosis. Suppurative nodule or something deeper?

Abstract: The authors report the case of a 62-year-old man with a history of total left hip arthroplasty nine years previously presenting with a large infiltrated plaque on the posterior area of the left thigh with three months of evolution without systemic symptoms or elevated inflammatory markers. Computed tomography of the left lower limb revealed a 12-centimeter linear extension of the lesion to the posterior part of the left proximal femur. Prosthesis joint infection, although rare, is a surgical complication to be taken into account, even if the surgery was performed many years before.

[Diagnosis and treatment of suppurative mediastinitis - a special chapter in the history of surgery (in Russian only)]. / Diagnostika i lechenie gnoinogo mediastinita - osobaia glava v istorii khirurgii.

It was reviewed a history of diagnosis and treatment of suppurative mediastinitis from ancient times to our time depending on inflammation type, localization and clinical features. An important role of national surgical school in the development of surgical treatment was emphasized.