ABSTRACT
A European multicenter study was performed to evaluate the performance of a new method, based on the transcription-reverse transcription concerted reaction (TRC-2), which enabled one-step amplification and real-time detection of the Mycobacterium tuberculosis 16S rRNA target directly in clinical specimens. A total of 633 respiratory and nonrespiratory specimens were tested, and the results were compared with those from smears and cultures. A total of 129 patients (Paris center) were followed up in order to evaluate the clinical performance of TRC-2. By using M. tuberculosis complex strains to inoculate sterile sputa, the detection limit of TRC-2 was found to be 30 to 50 CFU/ml. A total of 548 respiratory specimens and 59 extrapulmonary specimens were assessable. For pulmonary specimens, the sensitivities of TRC-2 and acid-fast smear were 86.8% and 50.4%, respectively (P = 0.002). The specificities were 97.5% and 100%, respectively. For extrapulmonary specimens, the sensitivities of TRC-2 and acid-fast smear were 83.3% and 8.3% (P < 0.0001), and the specificities were 95.8% and 100%, respectively. Fifteen of 129 patients were diagnosed with pulmonary tuberculosis (TB). The sensitivities of culture and TRC-2 were 80% (12/15) and 86.7% (13/15) (P = 0.16), and the specificities were 100% and 93.9%, respectively. Based on an 11.6% incidence of TB in our population, the positive predictive values of TRC-2 and culture were 81.3% and 100%, respectively, and the negative predictive values were 98.2% and 97.4%, respectively. These results demonstrated that detection of M. tuberculosis complex in clinical specimens by TRC-2 with ready-to-use reagents was an efficient and rapid method for the diagnosis of pulmonary and extrapulmonary TB.
Subject(s)
Molecular Diagnostic Techniques/methods , Mycobacterium tuberculosis/isolation & purification , Reverse Transcription , Transcription, Genetic , Tuberculosis/diagnosis , Adult , Body Fluids/microbiology , Europe , Female , Humans , Male , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/growth & development , Predictive Value of Tests , RNA, Bacterial/genetics , RNA, Ribosomal, 16S/genetics , Sensitivity and Specificity , Time FactorsABSTRACT
On December 2006, an outbreak of gastroenteritis occurred at a residential-care facility for the elderly in northern Italy. Thirty-five of 61 individuals interviewed (attack rate, 57.4%) fell ill. In 94.3% of cases, the onset of illness was within 48 h of a Christmas party at the facility. Norovirus (NoV) was detected by RT-PCR in 24 of 31 individuals examined, including three asymptomatic food-handlers, in whom there was evidence of long-lasting excretion of viral particles. The identification of a sequence referring to the '2006a GII.4 NoV variant' in all examined strains supported the hypothesis of a common point source. This retrospective cohort study is the first report on an outbreak of NoV gastroenteritis in an Italian residential-care facility for the elderly.
Subject(s)
Caliciviridae Infections/epidemiology , Disease Outbreaks , Gastroenteritis/epidemiology , Norovirus/isolation & purification , Residential Facilities , Adult , Aged , Aged, 80 and over , Caliciviridae Infections/virology , Chi-Square Distribution , Gastroenteritis/virology , Humans , Middle Aged , Norovirus/genetics , Risk Factors , Statistics, Nonparametric , Viral Proteins/genetics , Virus SheddingABSTRACT
BACKGROUND: Combination therapy with pegylated interferon (peginterferon) plus ribavirin is associated with several side effects, including neutropenia and infection. AIMS: To evaluate the incidence of neutropenia and infection between all consecutive patients with hepatitis C who were treated in two centers with peginterferon-alfa-2a and peginterferon-alfa-2b, in combination with ribavirin and actively monitored for occurrence of any infection. METHODS: A total of 319 consecutive patients with chronic hepatitis C received once-weekly peginterferon alfa-2b at a weight-adjusted dose (n=162) or peginterferon alfa-2a at a flat dose (n=157), plus ribavirin. RESULTS: Neutropenia was observed in 53 patients overall (17%). There were 73 infections in 73 subjects (23% of the treated population); 4/73 required hospitalization. Infections included respiratory infections (n=23), cellulitis (n=17), dental abscesses (n=13), gastroenteric infections (n=2), and other types of infections (n=18). The incidence of all infections was significantly associated with age, especially over 60 years (p<0.01) but not with neutropenia or type of pegylated interferon. CONCLUSIONS: During the treatment with pegylated interferons and ribavirin, we did not find a correlation between neutropenia and infections. This result provides a support for the notion that current guidelines for pegylated interferons dose reduction in the treatment of chronic hepatitis C for hematologic toxicity could be overly strict.
Subject(s)
Antiviral Agents/adverse effects , Hepatitis C, Chronic/drug therapy , Infections/epidemiology , Interferon-alpha/adverse effects , Neutropenia/epidemiology , Polyethylene Glycols/adverse effects , Ribavirin/adverse effects , Adolescent , Adult , Antiviral Agents/administration & dosage , Antiviral Agents/therapeutic use , Drug Therapy, Combination , Female , Hepacivirus/drug effects , Hepatitis C, Chronic/virology , Humans , Incidence , Infections/etiology , Interferon alpha-2 , Interferon-alpha/administration & dosage , Interferon-alpha/therapeutic use , Leukocyte Count , Male , Middle Aged , Neutropenia/etiology , Neutrophils/cytology , Polyethylene Glycols/administration & dosage , Polyethylene Glycols/therapeutic use , Recombinant Proteins , Ribavirin/administration & dosage , Ribavirin/therapeutic use , Treatment OutcomeABSTRACT
This study was aimed at investigating the possible involvement of the actin cytoskeleton in the modulation of host permissiveness to A/NWS/33 human influenza virus infection in two mammalian (MDCK and LLC-MK2) cell lines in vitro. During the early stages of infection, no appreciable association between incoming NWS/33 virions and cortical actin was detectable in the permissive MDCK model by confocal microscopy, while extensive colocalization and a slower infection progression were observed in LLC-MK2 cells. In the latter model, we also demonstrated the inability of the virus to carry out multiple replication cycles, irrespective of the presence of cleaved HA subunits in the released virions. Treatment with the actin-depolymerizing agent cytochalasin D significantly increased the infection efficiency in LLC-MK2 cells, while a detrimental effect was observed in the MDCK cell line. Our data suggest a selective role of the actin network in inducing a restriction to influenza virus replication, mostly depending on its molecular organization, the host cell type and virus replication phase.
Subject(s)
Actins/metabolism , Cytoskeleton/virology , Influenza A virus/physiology , Virus Replication , Actins/antagonists & inhibitors , Animals , Cell Line , Cytochalasin D/pharmacology , Cytoskeleton/metabolism , Dogs , Macaca mulatta , Microscopy, ConfocalABSTRACT
INTRODUCTION: Colorectal cancer is still one of the many factors of death both in males and in females. To date, the most important prognostic factors are mainly related to the pathological stage of the disease. AIM OF THE STUDY: The purpose of this study was to analyze the possible role of tumor circumferential localization on the colonic wall (mesenteric (M) or antimesenteric (AM)) as a possible prognostic factor. In this study, we compare the localization of the tumor with patient's survival. The hypothesis of this study is that M tumors, closer to blood and lymphatic vessels, should be more aggressive in terms of hematogenous and lymphatic spread compared to the AM tumors. PATIENTS AND METHODS: All patients undergoing curative resection for colorectal cancer were enrolled in this study; there was no statistical difference for age, sex and co-morbidity. The histopathological examination was carried out in the standard manner. Next, we have taken care to survival of neoplastic patients by examining of our 5-year follow-up archive: we divided patients in different groups concerning the different tumor stage and we compare these results with the different localizations of tumor at the operation. RESULTS: In 45% of cases, we were able to distinguish the different localizations M (160 patients) or AM (47 patients) and this difference is statistically significant (P<0.0001, Pearson Chi-Square-test (PCS-t)). The number of metastatic nodes is statistically higher in the M group compared to the AM group one (P=0.003949). Medium time of follow-up was 36.54 months; AM and M patients have a rather similar survival, only at the end the two curves seem to change but not in a significant manner. Only if we consider the difference between the two groups comparing T3 tumor can we observe a statistically significant difference (P<0.005). CONCLUSIONS: In conclusion, the localization of M or AM colorectal cancer is feasible in 45% of cases. M tumors have significantly more lymph nodes metastases but a better 5-year survival than AM tumors. A possible explanation for such results might be the different pattern of diffusion of cancer cells.
Subject(s)
Colorectal Neoplasms/mortality , Colorectal Neoplasms/pathology , Lymph Nodes/surgery , Mesentery/surgery , Colorectal Neoplasms/therapy , Follow-Up Studies , Humans , Lymph Nodes/pathology , Lymphatic Metastasis , Mesentery/pathology , Neoadjuvant Therapy , PrognosisABSTRACT
Detection of Plasmodium ovale by use of a nested PCR assay with a novel Plasmodium ovale primer set was superior to detection of Plasmodium ovale by real-time PCR assays. Nested PCR was also better at detecting P. malariae. The detection of P. ovale in many patients first admitted >2 months following their return to Italy indicated that P. ovale relapses are common.
Subject(s)
Malaria/diagnosis , Malaria/parasitology , Plasmodium ovale/genetics , Plasmodium ovale/isolation & purification , Polymerase Chain Reaction/methods , Polymorphism, Genetic , Adult , Animals , Child , Child, Preschool , Humans , Middle Aged , RNA, Protozoan/genetics , RNA, Ribosomal/genetics , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
Rapid identification of porcine Brachyspira species is required in order to differentiate pathogenic from non-pathogenic species. The aim of our study was to compare a recently described genetic method based on polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP), nox RFLP-PCR assay, and three species-specific PCRs described previously in the literature with a 16S rRNA gene RFLP-PCR discriminatory reference assay (16S RFLP-PCR) for the identification of Brachyspira spp. of swine origin. In this study, 20 porcine spirochaetal strains were identified and compared to 33 reference strains by 16S RFLP-PCR and nox RFLP-PCR and three species-specific PCRs. RFLP-PCR methods showed concordant results for 47 strains and discordances for 6 strains (2 differently identified and 4 not revealed by nox RFLP-PCR). In our hands species-specific PCRs showed concordant results with 16S and nox RFLP-PCR for 43 strains and discordances for 10 strains (2 differently identified and 8 not amplified). The same results observed testing the 20 field-isolated spirochaetes were obtained for the corresponding porcine faecal samples. The detection limit was 10(2) -10(3) cells/g of faeces for 16S rRNA gene PCR and 10(4) cells/g of faeces for nox PCR. In our experience nox RFLP-PCR appeared successful for the speciation of B. hyodysenteriae reserving 16S RFLP-PCR for all other pathogenic and non-pathogenic Brachyspira species. Among the species-specific PCR assays tested only that for B. pilosicoli was useful in our hands.
Subject(s)
Intestinal Diseases/veterinary , Polymorphism, Restriction Fragment Length , Spirochaetales Infections/veterinary , Spirochaetales/genetics , Swine Diseases/diagnosis , Swine Diseases/microbiology , Animals , Base Sequence , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Feces/microbiology , Intestinal Diseases/diagnosis , Intestinal Diseases/microbiology , NADPH Oxidases/genetics , Phylogeny , Polymerase Chain Reaction/veterinary , RNA, Ribosomal, 16S/chemistry , RNA, Ribosomal, 16S/genetics , Species Specificity , Spirochaetales/classification , Spirochaetales/enzymology , Spirochaetales/isolation & purification , Spirochaetales Infections/diagnosis , Spirochaetales Infections/microbiology , SwineABSTRACT
The aim of this study was to compare and evaluate the time required to isolate Brachyspira hyodysenteriae and Brachyspira pilosicoli from porcine faeces. This was done using previously described selective media (spectinomycin) S400, (colistin, vancomycin and spectinomycin) CVS and (spectinomycin, vancomycin, colistin, spiramycin and rifampin with swine faecal extract) BJ, compared with the method based on blood agar modified medium, with spectinomycin and rifampin (BAM-SR), including a pre-treatment step. Fourteen spirochaetal strains were obtained in pure cultures after 5 days (48 h in BAM-SR primary plate and three passages every 24 h in brain heart infusion (BHI) without antibiotics) pre-treating simulated samples in brain heart infusion broth with spectinomycin (400 microg/ml) and rifampin (15 microg/ml), before streaking on the selective BAM-SR medium. Spirochaetes from samples in S400, CVS and BJ, with and without pre-treatment, were obtained in pure cultures only after repeatedly transferring on plates of the same selective medium requiring 15-18 days according to the strain. BAM-SR used after the pre-treatment step showed a detection limit ranging from 3.5 x 10(2) to 6.7 x 10(7) cells/g faeces and was the only method able to support the growth of spirochaetes after 48 h.
Subject(s)
Dysentery/veterinary , Spirochaetales Infections/veterinary , Spirochaetales/isolation & purification , Swine Diseases/diagnosis , Animals , Colony Count, Microbial/veterinary , Culture Media/chemistry , Dysentery/diagnosis , Dysentery/microbiology , Feces/microbiology , Rifampin , Spectinomycin , Spirochaetales Infections/diagnosis , Spirochaetales Infections/microbiology , Swine , Swine Diseases/microbiology , Time FactorsABSTRACT
AIM: Purpose of the study was to evaluate if the circumferential location of colorectal cancer may be identified as a possible prognostic factor. The hypothesis is that tumours located on the antimesenteric (AM) side could have a better prognosis than tumours located on the mesenteric (M) side. METHODS: All patients undergoing curative resection for colorectal cancer were enrolled in the study. The specimens were sent to the pathologist to define the exact location of the tumour, the histological type, grading, T, N status as well as lymphatic, vascular and neural invasion, peritumoural lymphoid reaction, desmoplasia and microsatellite instability. Statistical analyses were performed using the test for proportions (with continuity correction), the Pearson Chi-square test and generalised linear models; p<0.05 were considered statistically significant. RESULTS: From August 2000 to August 2002, 255 patients were enrolled in the study. There was a significantly higher incidence of tumours located on the M (101) compared with the AM (37) site (p<0.0001). M located tumours were associated with higher numbers of metastatic lymph nodes (N1 and N2; p-value=0.014), whereas AM tumours were associated with involved lymph nodes in only 5/37 (13.5%) of tumours. There was no statistically significant relation between AM versus M location and T status: the Pearson Chi-Square test showed that the lymph node involvement and the location (M versus AM) are not statistically independent variables (p-value=0.014). CONCLUSIONS: Our preliminary results show that when M or AM tumour identification is possible, tumour location can be regarded as a prognostic factor. Further longer studies on recurrence rate and survival are required to validate these findings and the clinical usefulness of this putative prognostic factor.
Subject(s)
Colorectal Neoplasms/pathology , Lymph Nodes/pathology , Adult , Aged , Aged, 80 and over , Colorectal Neoplasms/surgery , Female , Humans , Lymphatic Metastasis , Male , Mesentery/pathology , Middle Aged , Neoplasm Recurrence, Local , PrognosisABSTRACT
A new molecular diagnostic method "Malaria-IBRIDOGEN" (Amplimedical S.p.A.--Bioline Division, Turin, Italy) based on a plate-hybridization assay for the simultaneous detection and identification of human malaria parasites was evaluated in this study. A target DNA sequence of the plasmodial 18S ribosomal RNA gene was amplified by polymerase chain reaction (PCR) and hybridized in microtiter wells with five biotinylated probes each specific for Plasmodium falciparum, P. vivax, P. malariae, P. ovale and the beta-globine human gene, respectively. Compared to the nested-PCR actually used in our laboratory for the molecular diagnosis of malaria, "Malaria-IBRIDOGEN" revealed an overall sensitivity of 100% (51/51) for the four human Plasmodium species testing 100 whole blood samples from people with malaria-like symptoms and fever. Specificity was 92% (45/49) considering four discordant samples as "false positive" by "Malaria-IBRIDOGEN". The assay showed a threshold of parasite density (detection limit) of 0.07 P. falciparum parasites/microliter, 0.15-1.5 P. vivax parasites/microliter, 0.3 P. malariae parasites/microliter and 0.4 P. ovale parasites/microliter of whole blood, respectively. This assay could be successfully applied to the laboratory diagnosis of malaria as a useful aid to microscopy.
Subject(s)
Malaria, Falciparum/diagnosis , Nucleic Acid Hybridization/methods , Plasmodium falciparum/genetics , Plasmodium falciparum/isolation & purification , Animals , Electrophoresis, Agar Gel , Evaluation Studies as Topic , Humans , Malaria, Vivax/diagnosis , Plasmodium malariae/genetics , Plasmodium malariae/isolation & purification , Plasmodium ovale/genetics , Plasmodium ovale/isolation & purification , Plasmodium vivax/genetics , Plasmodium vivax/isolation & purification , Polymerase Chain Reaction , Sensitivity and SpecificityABSTRACT
A TaqMan-based real-time PCR qualitative assay for the detection of three species of malaria parasites-Plasmodium falciparum, P. ovale, and P. vivax-was devised and evaluated using 122 whole-blood samples from patients who had traveled to areas where malaria is endemic and who presented with malaria-like symptoms and fever. The assay was compared to conventional microscopy and to an established nested-PCR assay. The specificity of the new assay was confirmed by sequencing the PCR products from all the positive samples and by the lack of cross-reactivity with Toxoplasma gondii and Leishmania infantum DNA. Real-time PCR assay showed a detection limit (analytical sensitivity) of 0.7, 4, and 1.5 parasites/ micro l for P. falciparum, P. vivax, and P. ovale, respectively. Real-time PCR, like nested PCR, brought to light errors in the species identification by microscopic examination and revealed the presence of mixed infections (P. falciparum plus P. ovale). Real-time PCR can yield results within 2 h, does not require post-PCR processing, reduces sample handling, and minimizes the risks of contamination. The assay can therefore be easily implemented in routine diagnostic malaria tests. Future studies are warranted to investigate the clinical value of this technique.
Subject(s)
Malaria/diagnosis , Plasmodium falciparum/isolation & purification , Plasmodium ovale/isolation & purification , Plasmodium vivax/isolation & purification , Polymerase Chain Reaction/methods , Animals , Base Sequence , Computer Systems , Cross Reactions , DNA Primers , DNA, Ribosomal/genetics , Diagnostic Tests, Routine , Genome, Protozoan , Humans , Leishmania infantum/isolation & purification , Plasmodium falciparum/genetics , Plasmodium ovale/genetics , Plasmodium vivax/genetics , RNA, Ribosomal, 18S/genetics , Sensitivity and Specificity , Toxoplasma/isolation & purificationABSTRACT
The cellular distribution of the human cytomegalovirus (HCMV)-specific UL83 phosphoprotein (pp65) and UL123 immediate-early protein (IEp72) in lytically infected human embryo fibroblasts was studied by means of indirect immunofluorescence and confocal microscopy. Both proteins were found to have a nuclear localization, but they were concentrated in different compartments within the nuclei. The pp65 was located predominantly in the nucleoli; this was already evident with the parental viral protein, which was targeted to the above nuclear compartment very soon after infection. The nucleolar localization of pp65 was also observed at later stages of the HCMV infectious cycle. After chromatin extraction (in the so-called in situ nuclear matrices), a significant portion of the pp65 remained associated with nucleoli within the first hour after infection, then gradually redistributed in a perinucleolar area, as well as throughout the nucleus, with a granular pattern. A quite different distribution was observed for IEp72 at very early stages after infection of human embryo fibroblasts with HCMV; indeed, this viral protein was found in bright foci, clearly observable in both non-extracted nuclei and in nuclear matrices. At later stages of infection, IEp72 became almost homogeneously distributed within the whole nucleus, while the foci increased in size and were more evenly spread; in several infected cells some of them lay within nucleoli. This peculiar nuclear distribution of IEp72 was preserved in nuclear matrices as well. The entire set of data is discussed in terms of the necessity of integration for HCMV-specific products into the pre-existing nuclear architecture, with the possibility of subsequent adaptation of nuclear compartments to fit the needs of the HCMV replicative cycle.
Subject(s)
Cell Nucleus/metabolism , Cell Nucleus/virology , Fibroblasts/metabolism , Fibroblasts/virology , Immediate-Early Proteins/metabolism , Nuclear Matrix/metabolism , Phosphoproteins/metabolism , Viral Matrix Proteins/metabolism , Viral Proteins/metabolism , Cell Fractionation , Cell Nucleolus/metabolism , Cell Nucleus/ultrastructure , Cells, Cultured , Cytomegalovirus/physiology , Embryo, Mammalian/cytology , Fibroblasts/ultrastructure , Fluorescent Antibody Technique, Indirect , Humans , Immediate-Early Proteins/ultrastructure , Lung/metabolism , Lung/ultrastructure , Lung/virology , Microscopy, Confocal , Nuclear Matrix/ultrastructure , Nuclear Matrix/virology , Phosphoproteins/ultrastructure , Subcellular Fractions , Viral Matrix Proteins/ultrastructure , Viral Proteins/ultrastructure , Virus ReplicationABSTRACT
AIM: We evaluated the usefulness of (99m)Tc-tetrofosmin axillary pinhole (P)-SPECT in breast cancer (BC) non palpable axillary lymph node metastasis detection compared with conventional planar and SPECT scintimammography. METHODS: We studied prospectively 188 consecutive patients with suspected primary BC, negative at axillary clinical examination. Ten minutes after 740 MBq (99m)Tc-tetrofosmin injection, planar and SPECT scintimammography were acquired, followed by axillary P-SPECT imaging. RESULTS: At histology, 12 patients had benign mammary lesions and 176 had BC. Axillary lymph node dissection (ALND) was performed in all BC patients, bilaterally in 3 cases: 74/179 axillae had metastases. P-SPECT showed a significantly higher overall sensitivity than SPECT and planar (93.2% vs 85.1% and 36.5%, respectively; p<0.05 and p<0.0005, respectively) and was false negative in 5 patients with 1 metastatic node each, micrometastatic in 4/5 cases; SPECT and planar were also false negative in these 5 cases and in 6 and in 42 further cases, respectively. P-SPECT added important prognostic information by distinguishing single from multiple and pound 3 from >3 nodes; only P-SPECT defined the exact number of nodes in 15/25 patients with 2-4 nodes. P-SPECT showed the highest accuracy and NPV: 92.7% and 95%, respectively (SPECT 90.5% and 90%, respectively; planar 73.2% and 68.9%, respectively). CONCLUSION: (99m)Tc-tetrofosmin axillary P-SPECT appears highly accurate in BC non palpable axillary lymph node metastasis detection and significantly more sensitive than both planar and SPECT, its few false negative results predominantly concerning micrometastases; moreover, only P-SPECT gave additional important prognostic information. Given its very high NPV, P-SPECT could also be used to better select patients who might avoid ALND.
Subject(s)
Breast Neoplasms/diagnostic imaging , Breast Neoplasms/pathology , Lymph Nodes/diagnostic imaging , Organophosphorus Compounds , Organotechnetium Compounds , Tomography, Emission-Computed, Single-Photon/methods , Adult , Aged , Axilla , Breast Neoplasms/diagnosis , Female , Humans , Lymphatic Metastasis , Male , Middle Aged , Palpation , Radiopharmaceuticals , Reproducibility of Results , Sensitivity and Specificity , Tomography, Emission-Computed, Single-Photon/instrumentationABSTRACT
Susceptibility of Mycobacterium bovis strains to antituberculous drugs (isoniazid and rifampin) was detected by radiometric BACTEC 460TB system. M.bovis strains were isolated from tissue samples showing tuberculous lesions collected at an abbattoir from cattle belonging to 47 tuberculosis outbreaks occurring in Northern Italy in 1995-1999. Forty-six out of 61 strains (75.4%) resulted susceptible to both isoniazid and rifampin. Thirteen strains (21.3%) were resistant to isoniazid only. No strains showed resistance to rifampin only. Two strains (3.3%) resulted resistant to both drugs, showing antituberculous multidrug-resistance. Given the compulsory eradication program of bovine tuberculosis by elimination of infected animals and the ban on antituberculous drug treatments in animals, detection of resistant M. bovis strains appears of great interest.
Subject(s)
Antitubercular Agents/pharmacology , Isoniazid/pharmacology , Microbial Sensitivity Tests/methods , Mycobacterium bovis/drug effects , Radiometry/methods , Rifampin/pharmacology , Tuberculosis, Bovine/drug therapy , Animals , Antitubercular Agents/therapeutic use , Cattle , Isoniazid/therapeutic use , Mycobacterium bovis/isolation & purification , Reproducibility of Results , Rifampin/therapeutic use , Tuberculosis, Bovine/diagnosis , Tuberculosis, Bovine/microbiologyABSTRACT
The species-specific nested-PCR previously described by Snounou and others for detecting the four parasite species that cause human malaria is evaluated in the current study testing 230 blood samples. The results are compared with those obtained by microscopy and, for 101 samples out of 230, with those previously obtained by a genus-specific PCR based method (pg-PCR) followed by species-specific Southern-blot hybridization. All blood specimens were obtained from patients (127 foreigners and 103 Italians) with a suspect clinical diagnosis of imported malaria in Italy: 76 were positive by microscopy and 83 were positive by nested-PCR. The last method also revealed 10 double infections (8 foreigners and 2 Italians) which were not identified by microscopy or by pg-PCR with species-specific Southern-blot hybridization. Fifty-four out of 83 positive samples tested by nested-PCR were submitted to genomic sequence analysis, which confirmed the presence of DNA region portion encoding the 18S rRNA corresponding to the Plasmodium species identified by nested-PCR. These results demonstrate that the nested-PCR assay surpasses microscopy and pg-PCR with species-specific Southern-blot hybridization, both in sensitivity and in diagnostic accuracy. Moreover, it is quicker because it requires no further blotting or hybridization of PCR amplification products. This method also offers a clear advantage in the detection of mixed infections, which is important not only for successful medical treatment but also for the study of malaria epidemiology. Finally, our study also highlights the value of genomic sequence analysis for validating PCR results.
Subject(s)
Malaria/parasitology , Plasmodium/classification , Animals , DNA, Protozoan/chemistry , DNA, Protozoan/genetics , Humans , Italy , Malaria/blood , Malaria/diagnosis , Plasmodium/genetics , Plasmodium/isolation & purification , Polymerase Chain Reaction/methods , RNA, Ribosomal, 18S/chemistry , RNA, Ribosomal, 18S/genetics , Retrospective Studies , Sensitivity and Specificity , Sequence Alignment , Sequence Analysis, DNAABSTRACT
PURPOSE: To assess the usefulness of stereotactic vacuum-assisted core breast biopsy (VCBB) performed using a stereotactic add-on device and film-screen technology with the patient in an upright seated position. MATERIAL AND METHODS: We reviewed a series of 129 women with non-palpable mammographic abnormalities who required stereotactic VCBB from December 1999 to November 2000. Twenty-seven (20.9%) cases were excluded due to difficulties in keeping the correct position during the procedure, while the other 102 (79.1%) underwent successful VCBB. Patients with lesions consisting of either atypical ductal hyperplasia or lobular carcinoma in situ were considered for excisional biopsy. Patients with either ductal carcinoma in situ or infiltrating breast cancer were referred for definitive surgery. The results of stereotactic VCBB were correlated to the subsequent surgical histology. RESULTS: Stereotactic VCBB was interrupted because of bleeding in 1 case and vasovagal reaction in 5 cases. Two haematomas occurred after the procedure. Overall underestimation rate was 10.5%. No new lesions were discovered after a mean follow-up of 18.7 months. CONCLUSION: Stereotactic VCBB performed using a standard add-on device with the patient in an upright seated position and analog technology is feasible in about 80% of cases, has a low complication rate, is not significantly time-consuming, and can offer the same accuracy as dedicated prone equipment.
Subject(s)
Biopsy, Needle/methods , Breast/pathology , Stereotaxic Techniques , Biopsy, Needle/adverse effects , Breast Neoplasms/diagnosis , Female , Humans , Mammography , Posture , Radiography, Interventional , Retrospective Studies , Stereotaxic Techniques/adverse effects , VacuumABSTRACT
Brachyspira (Serpulina) pilosicoli of human origin interfere with the growth of Clostridium perfringens alpha-toxin producer reducing the clostridial growth area and colonies number when bacteria were cultivated together in sheep blood agar plates. The growth inhibition of C. perfringens was only observed when B. (S.) pilosicoli grew 72-96 hours sooner than C. perfringens and after the inoculum of this latter the plates were anaerobically incubated for additional 48 hours. The phenomenon was observed at concentrations of B. (S.) pilosicoli ranging from 10(7) to 10(4) CFU/ml and at concentrations of C. perfringens ranging from 10(7) to 10(1) CFU/ml when the bacteria were 0-10 mm away from each other. When B. (S.) pilosicoli and C. perfringens were inoculated at the same time and when B. (S.) pilosicoli grew 24-48 hours sooner than C. perfringens, the clostridial growth inhibition was not appreciated and only a cooperative haemolysis was observed between the bacteria.
Subject(s)
Antibiosis/physiology , Bacterial Toxins/biosynthesis , Clostridium perfringens/physiology , Spirochaetales/physiology , Bacterial Toxins/antagonists & inhibitors , Clostridium perfringens/growth & development , Hemolysis , Humans , Microbial Sensitivity Tests , Spirochaetales/growth & developmentABSTRACT
Brachyspira (Serpulina) pilosicoli related to intestinal spirochaetosis were found to interfere in vitro with the haemolytic activity and the growth of Staphylococcus aureus beta-toxin producer. This interference was clearly appreciated because a reduction of the zone of the staphylococcal beta-toxin activity, the reduction and/or absence of cooperative haemolysis between bacteria, and the growth reduction of S. aureus were observed when B. (S.) pilosicoli were grown 72-96 hours sooner than S. aureus and after the inoculum of the latter the plates were anaerobically incubated for additional 48-72 hours. The phenomenon was more clearly observed when B. (S.) pilosicoli had a concentration of 8x10(6)-8x10(7) CFU/ml and S. aureus at a concentration ranging from 10(7) to 10(1) CFU/ml was inoculated at a distance from the streaks of B. (S.) pilosicoli ranging from 0-10 mm. When B. (S.) pilosicoli and S. aureus were inoculated at the same time and when B. (S.) pilosicoli grew 24-48 hours sooner than S. aureus only a cooperative haemolysis was observed.
Subject(s)
Antibiosis/physiology , Bacterial Toxins/biosynthesis , Hemolysis , Spirochaetales/physiology , Staphylococcus aureus/physiology , Bacterial Toxins/antagonists & inhibitors , Humans , Microbial Sensitivity Tests , Spirochaetales/growth & development , Staphylococcus aureus/growth & developmentABSTRACT
The aim of this study was to evaluate the usefulness of (99m)Tc-tetrofosmin single-photon emission tomography (SPET) in the detection of both primary breast cancer and axillary lymph node metastasis. We studied 192 consecutive patients in whom primary breast cancer was suspected on the basis of mammography and/or physical examination. After intravenous injection of 740 MBq (99m)Tc-tetrofosmin, both planar and SPET scintimammography was performed in all patients using a rectangular dual-head gamma camera equipped with low-energy, high-resolution, parallel-hole collimators. In 175 patients with breast cancer at histology, the per-lesion overall sensitivity of SPET and planar imaging for the detection of breast cancer was 95.8% and 75.9% (P<0.0005), respectively. The sensitivity of SPET and planar imaging was, respectively, 96.5% and 79.5% in palpable (P<0.0005) and 90% and 45% in non-palpable lesions (P<0.01). With regard to lesion size, the sensitivity of SPET and planar imaging was, respectively, 90.5% and 45.2% in lesions < or =10 mm ( P<0.0005), 95.3% and 81.4% in lesions of 11-20 mm (P<0.005), 100% and 84.6% in lesions of 21-30 mm (P<0.05) and 100% and 95.8% in lesions >30 mm (P>0.05). In the remaining 17 patients with benign mammary lesions at histology, per-lesion overall specificity of SPET and planar imaging was 76.2% and 85.7% (P>0.05), respectively. Neither SPET nor planar imaging showed false-positive results in non-palpable lesions or in those < or =10 mm. In 173 breast cancer patients submitted to axillary lymph node dissection (ALND), per-axilla overall sensitivity of SPET and planar imaging in the detection of axillary lymph node metastasis was 93% and 52.3% ( P<0.0005), respectively. The sensitivity of SPET and planar imaging was, respectively, 100% and 82.6% in palpable nodes (P>0.05), 90.5% and 41.3% in non-palpable nodes (P<0.0005), 92.8% and 35.7% in the presence of < or =3 nodes ( P<0.0005) and 93.2% and 68.2% in the presence of >3 nodes (P<0.005). The specificity of SPET and planar imaging was 91% and 100% (P<0.05), respectively. (99m)Tc-tetrofosmin SPET appears to be a reliable method for the detection of both primary BC and axillary lymph node metastasis, and its diagnostic accuracy exceeds that of (99m)Tc-tetrofosmin planar scintimammography. The use of SPET is particularly important in the identification of small non-palpable primary carcinomas and metastatic axillae with < or =3 non-palpable lymph nodes. More extensive use of SPET appears warranted in the management of breast cancer patients.