Sepsis is a common and lethal syndrome: although outcomes have improved, mortality remains high. No specific anti-sepsis treatments exist; as such, management of patients relies mainly on early recognition allowing correct therapeutic measures to be started rapidly, including administration of appropriate antibiotics, source control measures when necessary, and resuscitation with intravenous fluids and vasoactive drugs when needed. Although substantial developments have been made in the understanding of the basic pathogenesis of sepsis and the complex interplay of host, pathogen, and environment that affect the incidence and course of the disease, sepsis has stubbornly resisted all efforts to successfully develop and then deploy new and improved treatments. Existing models of clinical research seem increasingly unlikely to produce new therapies that will result in a step change in clinical outcomes. In this Commission, we set out our understanding of the clinical epidemiology and management of sepsis and then ask how the present approaches might be challenged to develop a new roadmap for future research.

https://www.thelancet.com/pdfs/journals/laninf/PIIS1473-3099(15)70112-X.pdf

Sepsis: a roadmap for future research

SUMMARY Sepsis is among the most common causes of death in hospitals. It arises from the host response to infection. Currently, diagnosis relies on nonspecific physiological criteria and culture-based pathogen detection. This results in diagnostic uncertainty, therapeutic delays, the mis- and overuse of antibiotics, and the failure to identify patients who might benefit from immunomodulatory therapies. There is a need for new sepsis biomarkers that can aid in therapeutic decision making and add information about screening, diagnosis, risk stratification, and monitoring of the response to therapy. The host response involves hundreds of mediators and single molecules, many of which have been proposed as biomarkers. It is, however, unlikely that one single biomarker is able to satisfy all the needs and expectations for sepsis research and management. Among biomarkers that are measurable by assays approved for clinical use, procalcitonin (PCT) has shown some usefulness as an infection marker and for antibiotic stewardship. Other possible new approaches consist of molecular strategies to improve pathogen detection and molecular diagnostics and prognostics based on transcriptomic, proteomic, or metabolic profiling. Novel approaches to sepsis promise to transform sepsis from a physiologic syndrome into a group of distinct biochemical disorders and help in the development of better diagnostic tools and effective adjunctive sepsis therapies.

New Approaches to Sepsis: Molecular Diagnostics and Biomarkers

Background
Blood culture is viewed as the golden standard for the diagnosis of sepsis but suffers from low sensitivity and long turnaround time. LightCycler SeptiFast (LC-SF) is a real-time multiplex polymerase chain reaction test able to detect 25 common pathogens responsible for bloodstream infections within hours. We aim to assess the accuracy of LC-SF by systematically reviewing the published studies.

/pdf/multiplex-pcr-system-for-rapid-detection-of-pathogens-in-4m84jvthl4.pdf

Multiplex PCR System for Rapid Detection of Pathogens in Patients with Presumed Sepsis – A Systemic Review and Meta-Analysis

Malaria is frequently imported into nonendemic industrialized areas. In this study we collated data on the reported malaria cases in industrialized countries during the period 1985-95, with the object of identifying trends and promising strategies. The main outcome measures were incidence, case-fatality rates (CFRs), and attack rates in tourists returning from Kenya. Our survey showed gross underreporting and marked heterogeneity in the type and availability of national data. The total incidence or reported numbers of malaria infections in Europe increased from 6840 in 1985 to 7244 in 1995, with a peak of 8438 in 1989. The principal importing countries were France, Germany, Italy, and the United Kingdom. In the former USSR, the reported annual incidence dropped from 1145 in 1989 to 356 in 1990 after cessation of activities in Afghanistan. Among the imported species of malaria parasite, Plasmodium falciparum was identified in an increasing proportion, the CFR ranging from 0% to 3.6%, with consistently high rates in Germany. The attack rates among travellers to Kenya in 1990-95 were high, ranging from 18 to 207 per 100,000 travellers. Our findings indicate that in industrialized countries malaria is associated with a high CFR and remains a public health problem. Irregular surveillance and lack of homogeneity in the collected data hinder the assessment of incidences, risk groups, and the efficacy of chemoprophylaxis.

https://www.who.int/bulletin/archives/77(7)560.pdf

Imported malaria (1985-95): trends and perspectives.

By shortening the time to pathogen identification and allowing for detection of organisms missed by blood culture, new molecular methods may provide clinical benefits for the management of patients with sepsis. While a number of reviews on the diagnosis of sepsis have recently been published we here present up-to-date new developments including multiplex PCR, mass spectrometry and array techniques. We focus on those techniques that are commercially available and for which clinical studies have been performed and published.

Molecular diagnosis of sepsis: New aspects and recent developments

OBJECTIVES: To evaluate the potential improvement of antimicrobial treatment by utilizing a new multiplex polymerase chain reaction (PCR) assay that identifies sepsis-relevant microorganisms in blood. DESIGN: Prospective, observational international multicentered trial. SETTING: University hospitals in Germany (n = 2), Spain (n = 1), and the United States (n = 1), and one Italian tertiary general hospital. PATIENTS: 436 sepsis patients with 467 episodes of antimicrobial treatment. METHODS: Whole blood for PCR and blood culture (BC) analysis was sampled independently for each episode. The potential impact of reporting microorganisms by PCR on adequacy and timeliness of antimicrobial therapy was analyzed. The number of gainable days on early adequate antimicrobial treatment attributable to PCR findings was assessed. MEASUREMENTS AND MAIN RESULTS: Sepsis criteria, days on antimicrobial therapy, antimicrobial substances administered, and microorganisms identified by PCR and BC susceptibility tests. RESULTS: BC diagnosed 117 clinically relevant microorganisms; PCR identified 154. Ninety-nine episodes were BC positive (BC+); 131 episodes were PCR positive (PCR+). Overall, 127.8 days of clinically inadequate empirical antibiotic treatment in the 99 BC+ episodes were observed. Utilization of PCR-aided diagnostics calculates to a potential reduction of 106.5 clinically inadequate treatment days. The ratio of gainable early adequate treatment days to number of PCR tests done is 22.8 days/100 tests overall (confidence interval 15-31) and 36.4 days/100 tests in the intensive care and surgical ward populations (confidence interval 22-51). CONCLUSIONS: Rapid PCR identification of microorganisms may contribute to a reduction of early inadequate antibiotic treatment in sepsis.

Potential clinical utility of polymerase chain reaction in microbiological testing for sepsis.

Gene amplification using 16S rDNA primers has been proposed as a strategy for the diagnosis of bacterial meningitis. The aim of this study was to evaluate the performance of the MicroSeq 500 16S ribosomal DNA test (Applied Biosystems) from patients with suspected bacterial meningitis and CSF negative-culture in comparison to traditional methods. Twelve purulent culture-negative CSF samples were collected between January 2005 and January 2007. For DNA extraction, 500 microl of CSF samples were treated using the QIAamp mini kit (QIAGEN). The extracted DNA was examined amplifying 500 bp at the 5' end of 16S rRNA gene using MicroSeq500 16S rDNA Bacterial Identification PCR kit and the sequencing reactions were performed with the MicroSeq500 16S rDNA Bacterial Identification Sequencing kit (Applied Biosystems). The sequences were compared with those available in GenBank. For the culture-negative CSF samples the MicroSeq 500 16S rDNA yielded a positive result in 9 cases (75.0%): three samples were identified as Streptococcus. pneumoniae, three as Neisseria meningitidis, and the remaining 3 as Haemophilus influenzae, Abiotrophia defectiva and Porphyromonas gingivalis. The MicroSeq 500 16S ribosomal DNA test may improve the microbiological diagnosis of bacterial meningitis, especially when spinal fluid samples are obtained after the administration of antimicrobial therapy.

Evaluation of the MicroSeq 500 16S rDNA-based gene sequencing for the diagnosis of culture-negative bacterial meningitis.

Background: Malaria infections have become an increasing public health problem in Europe, especially those imported into nonendemic areas. The transmission and diffusion of malaria has increased, especially over the last decade, due to changes in agricultural practices, vector resistance to insecticides, and most relevantly, increasing international travel and the resistance of these parasites to chemophrophylaxis. This study investigates the epidemiologic factors of imported malaria in an area of Italy, as related to international travel and prophylaxis by Italian immigrants who have revisited their country of origin.



Method: All cases (175) of imported malaria detected at the Laboratory of Microbiology of Ospedali Riuniti in Bergamo, Italy, between 1984 and 1993 were studied epidemiologically for the following variables: age, sex, and nationality; travel destination, length of stay, and date of return; and pathogen(s) detected, chemoprophylaxis used, and clinical symptoms exhibited.



Results: A high prevalence of Plasmodium falciparum was detected in more than three quarters of the cases with 91.4% of these travelers having visited African countries. Only two subjects had received adequate, correct prophylaxis. Fever, headache, and fatigue were experienced most often; however, in a few cases, blood, exchange transfusion, or treatment for splenomegaly were required.



Conclusions: The results indicate that there is an emerging public health problem with immigrants who have resided in Italy for some time, revisited their country of origin, and consequently become infected with malaria, with specific prophylaxis not having been provided. This study emphasizes the importance of local epidemiologic studies, effective prophylaxis, and the need for those involved in the travel industry to promote specialized pretravel advice on a routine basis.

/pdf/ten-year-experience-with-imported-malaria-in-bergamo-italy-1apj7j42pt.pdf

Ten‐year Experience with Imported Malaria in Bergamo, Italy

Lymphadenitis can be caused by different gram positive and gram negative bacteria and non-tuberculous mycobacteria. Cervical lymphadenitis in children is thought to result from ingestion of or contact with environmental microrganisms. Chromobacterium violaceum is a common inhabitant of soil and water in tropical and sub tropical countries. In these parts of the world Chromobacterium violaceum is able to cause skin infection with diffuse pustular lesions and also multiple liver abscess with often fatal evolution in sepsis. We describe a case of cervical lymphadenitis caused by Chromobacterium violaceum in a 14-year-old boy, born in Guinea and resident in Italy for 7 years in a fair condition with general measurable swelling in the right lateral cervical region and with blood tests that showed increased inflammatory indices. The patient was subjected to surgical incision. Antibiotic therapy with ceftriaxone was continued for 10 days, then replaced successfully with oral ciprofloxacin on the basis of purulent material culture positive for Chromobacterium violaceum sensitive to fluoroquinolones. SUMMARY

Chromobacterium violaceum lymphadenitis successfully treated in a Northern Italian hospital.

Acute Schistosoma mansoni infection with progression to chronic lesion in italian travelers returning from Cameroon, West Africa: A diagnostic and prevention problem

Annibale Raglio

Papers

Potential clinical utility of polymerase chain reaction in microbiological testing for sepsis.

Evaluation of the MicroSeq 500 16S rDNA-based gene sequencing for the diagnosis of culture-negative bacterial meningitis.

Ten‐year Experience with Imported Malaria in Bergamo, Italy

Chromobacterium violaceum lymphadenitis successfully treated in a Northern Italian hospital.

Acute Schistosoma mansoni infection with progression to chronic lesion in italian travelers returning from Cameroon, West Africa: A diagnostic and prevention problem