Listeria monocytogenes infection in pregnancy and neonatal sepsis

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New method for rapid Susceptibility Testing on blood culture with HB&L system: preliminary data

Blood culture, although represents the gold standard in detecting the ethiological agent of sepsis, is rather rarely required in relation to the real diagnostic importance. The result of this test depends in fact on many factors (sample volume, time of collection, accuracy, antibiotic therapy, contamination, number of drawings, drawing site, interpretation difficulties, etc.) that are often considered by many clinicians so limited as to doubt about their actual value. The disadvantages are therefore represented by the lack of standardization but also by the low sensitivity and above all by the technical times too long for the clinical needs. Blood culture begins with the drawing of samples from the “septic” patient followed incubation of the bottles in automatic thermostated systems. In case of positive result (36 hours), the culture is Gram stained and streaked on solid media in order to obtain isolated colonies for the identification and the susceptibility testing (48 hours from positive result). The long time required for pathogen identification and susceptibility testing involves empirical broad spectrum antibiotic therapy that can promote the increase of bacterial resistance but also patient management costs. A clinically useful report should be available on short notice in order to guide the clinician to choose the most appropriate antibiotic. The microbiologist has therefore the hard work of reviewing the organization and the management of the procedures.We have therefore started to consider the possibility of treating the blood as an biological liquid in order to quickly determine the susceptibility of bacteria to antibiotics

La reazione polimerasica a catena (PCR) nella diagnosi delle infezioni genitali da Papillomavirus umano (HPV)

In a three year time (2001-2003) we evaluated the use of HPV testing and typing in 520 cytologically abnormal women (322 LSIL, 198 HSIL), in 25 squamous and 18 adeno invasive cancers. HPV testing was performed by polymerase chain reaction (PCR) using L1 consensus primers; HPV typing was performed using specific primers for low and high risk types. The prevalence of high risk HPV was: LSIL 41%; HSIL 74%; squamous invasive cancers 96% and adeno 100%. HPV 16 is the most represented type in all lesions. We also evaluated the usefulness of p53 typing in the early identification of women at risk for cervical cancer

Detection of Gardnerella vaginalis, Candida spp. and Trichomonas vaginalis DNA in symptomatic women

While vaginitis caused by Trichomonas vaginalis is now less frequent, fungal Candida spp. infections are frequently found and the bacterial vaginosis is one of the most common vaginal diseases caused by anaerobic microorganisms such as Gardnerella vaginalis. Purpose of this study is to evaluate the usefulness of a rapid molecular test for the diagnosis of vaginitis/bacterial vaginosis in symptomatic women. In our clinic, between January 2008 - June 2009, we admitted 1592 (388 were pregnant) symptomatic women with a specific request to test them for fungi, Trichomonas and Gardnerella on vaginal fluid. The samples were tested with the kit Affirm (Becton Dickinson) that provides results in 40 minutes and allows the simultaneous identification of the DNA of Gardnerella vaginalis, Candida spp. and Trichomonas vaginalis. One hundred and eight out of 388 pregnant women were positive only for Gardnerella; 53 for Candida and Gardnerella; 59 were positive only for Candida and 10 for Trichomonas. As to the remaining 1204 not pregnant patients, 356 were positive only for Gardnerella; 98 for Candida and Gardnerella, 143 were positive only for Candida and 21 for Trichomonas.A simultaneous positivity for Trichomonas and Candida or for Trichomonas and Gardnerella has not been observed in any case. Molecular testing is obviously more sensitive and specific than culture method and microscopic research, especially for the detection of Gardnerella. It also enables differential diagnosis between bacterial vaginosis and vaginitis and therefore allows a targeted therapeutic intervention

HPV DNA genotyping test in low and high grade intraephitelial lesions

The human papillomavirus (HPV) is certainly one of the few viruses associated with cancer and is present in a significant number of cancers of the anus, penis, vagina, vulva and in almost all cervical cancers.The aim of our work is to evaluate the impact of genotypes high-risk oncogenic (HR) in low-grade (LSIL) and high grade (HSIL) intraepithelial lesions. In it are also evaluated the impact of genotypes low oncogenic risk (LR) and the percentage of non-detection (NR) Papillomavirus DNA Between January 2008 and June 2009 we submitted for HPV DNA genotyping test 392 patients, seronegative for HIV1/2, of which 278 with LSIL and 114 with HSIL. In 250 patients tested positive for HPV DNA HR the type 16 is present in 102 patients (41%), the 18 in 62 (25%), the 31 in 37 patients (15%), the 33 in 13 patients (5%); also was limited the simultaneous presence of 16 and 18 (13 patients 5%) The search for HPV DNA is a modern method of study of cervical pathology. Optimizes the follow-up of intraepithelial lesions and the therapeutic intervention, avoiding the dangers of over/ undertreatment and allows to monitor women treated for cervical pathology

Polyoma BK virus infection in renal transplant recipients

Several studies reported a correlation between the human Polyomavirus BK (BKV) and interstitial nephritis (PVN: Polyoma Virus Nephropaty) in renal transplant recipients in whom immunosuppressive treatment is thought to facilitate or induce reactivation of the virus. In the present study we monthly evaluated the presence of BKV-DNA in plasma and urine of 29 kidney transplant recipients. We used a nested PCR for BKV-DNA screening in plasma and urine and a quantitative assay Real Time PCR in case of a positive screening result. The viral DNA has been detected in 48% of the patients samples: only in urine of six patients; in plasma of four and in both of two. Immunosuppressive therapy has been decreased in positive patients. The kidney loss occurred just in the two patients with high viral load in plasma and urine. The definitive diagnosis of BKV Nephropaty requires allograft biopsy though biomolecular test for BKV-DNA in urine (viruria) and in plasma (viremia) could be very important non-invasive method for the early diagnosis of PVN

Staphylococci with markers of antibiotic resistance collected from blood cultures

Introduction: Blood culture is still the gold standard for the detection of the causative agent of sepsis. Especially in intensive care patients and those with vascular catheters, the most common organisms isolated are coagulase-negative staphylococci (CoNS) and Staphylococcus aureus, both characterized by multidrug resistance. Purposes of our work are the study of the incidence of markers of resistance in staphylococci and evaluation of potential changes over the years. Materials and methods: In the period January 2008-June 2011 5239 blood cultures were analyzed.They were mainly obtained from the departments of Intensive Care, Cardiology, Hematology, General Medicine, Emergency Medicine, Infectious Diseases, Oncology, Pulmonology and Pediatric Hematoncology. The vials containing the blood were incubated in the BACTEC 9120 automated tool of Becton Dickinson and susceptibility testing performed with the Phoenix instrument of the same company. Results:Within a total of 5239 blood cultures, 3967 (75.7%) were negative and 1272 (24.3%) positive. Fungi were isolated in 6.2% (79) of the positive ones, Gram-negative bacteria in 24.6% (313) and Gram-positive bacteria in 69.2% (880). Within the latter, 187 (21.2%) were not staphylococcal isolates, 693 (78.8%) were stafiloccocci mainly represented by S. epidermidis, S. aureus, S. hominis, S. haemolyticus and S. saprophyticus. Of the 693 staphylococcal isolates, 436 (62.9%) were b lactamase producers, and between them 336 (77.1%) were methicillin resistant, while only 3 of 436 (0.69%) were S. aureus resistant to vancomycin as well.The incidence of markers of resistance was very high, especially in patients in intensive care and cardiac surgery, who are usually subjected to combined antibiotic therapy. In the three years studied there were no statistically significant differences in the resistance of staphylococci. Conclusions: The data show an alarming high number of multi-resistant staphylococci, which is often a real therapeutic challenge for the clinician. The interpretation of the meaning of the isolation of CoNS entails a thorough evaluation because they are commonly isolated from contaminated blood cultures

Endotoxin dosage in sepsis

Introduction. Endotoxin, a component of the cell wall of Gram-negative bacteria is a major contributor to the pathogenesis of septic shock and multiple organ failure (MOF). Its entry into the bloodstream stimulates monocytes/macrophages which once activated produce and release cytokines, nitric oxide and other mediators that induce systemic inflammation, endothelial damage, organ dysfunction, hypotension (shock) and MOF.The aim of this study is to evaluate the usefulness of a quantitative test for the dosage of endotoxin to determine the risk of severe Gram-negative sepsis. Materials and methods. In the period January 2009 - June 2011 we performed 897 tests for 765 patients, mostly coming from the emergency room and intensive care, of which 328 (43%) women (mean age 53) and 437 (57%) male (mean age 49). Fifty-nine patients, no statistically significant difference in sex, were monitored by an average of two determinations of EA.All patients had procalcitonin values significantly altered.The kit used was EAA (Endotoxin Activity Assay) Estor Company, Milan, which has three ranges of endotoxin activity (EA): low risk of sepsis if <0.40 units, medium if between 0.40 and 0.59; high if 0.60. Results. 78 out of 765 patients (10%) had a low risk, 447 (58%) a medium risk and 240 (32%) a high risk.The dosage of EA, combined with that of procalcitonin, has allowed a more targeted antibiotic therapy. Six patients in serious clinical conditions were treated by direct hemoperfusion with Toraymyxin, a device comprising a housing containing a fiber polypropylene and polystyrene with surface-bound polymyxin B, an antibiotic that removes bacterial endotoxins from the blood. Conclusions.The test is useful in risk stratification as well as Gram negative sepsis, to set and monitor targeted therapies, also based on the neutralization of endotoxin

Citomegalovirus: monitoraggio dell’infezione e della terapia nei pazienti trapiantati di rene

CMV infection is a major cause of disease following renal transplantation. Clinical diagnosis is difficult because the virus usually produces only few, if any, symptoms. Consequently rapid and sensitive diagnostic methods are needed, since clinically effective antiviral therapy is available. Qualitative-quantitative detection of CMV-DNA in leukocytes and in plasma and pp65-antigenaemia are the methods which allow to evidence viral replication activity. In this study we report the our experience about follow-up of 70 kidney transplant recipients. Results indicate that while active CMV infection occured in 25 patients (37,5%), only 11 patients (15,7%) showed antigenaemia and DNAemia values predictive of CMV disease. In this study we also evaluated the efficacy of pre-emptive therapy

Listeria monocytogenes infection in pregnancy and neonatal sepsis

Authors report a fatal neonatal sepsis caused by Listeria monocytogenes. While the diagnostic procedure aimed to identify the microrganism is described, it is emphasized the importance to recover Streptococcus agalactiae (GBS) and L. monocytogenes by means of vaginal-rectal swab culture. The intrapartum screening for L. monocytogenes, by Polymerase Chain Reaction (PCR) providing results in 75 minutes is also evaluated