Prevalence, clinical features and quantification of genital non-viral infections.

We conducted a study of the prevalence, clinical features and microscopy findings, by retrospective case-notes survey, of six non-viral organisms, among 1718 attendees at a genitourinary (GU) medicine clinic in England. An in-house assay for six non-viral infections was used and quantitation of ureaplasmas performed. The prevalences of the six organisms were: Chlamydia trachomatis (CT), 7.1%; Neisseria gonorrhoeae (NG), 0.6%; Mycoplasma genitalium (MG), 1.0%; Trichomonas vaginalis, 0.2%; Ureaplasma urealyticum, 16.1%; Ureaplasma parvum, 35.6%. Among men (but not women) there were significantly raised odds ratios compared with that for U. parvum, for the symptom of discharge with CT, 7.30; MG, 6.43; NG 19.29; dysuria with CT, 5.89 and MG, 5.95; and the microscopy finding of >4 pus cells per high power field with: CT, 7.22; MG, 4.58 and NG 22.31. Evaluation of a possible link between quantitation of U. urealyticum and urethritis did not confirm research findings elsewhere.

[Unusual presentation of hypertriglyceridemic acute pancreatitis in a child]. / Présentation inhabituelle d'une pancréatite aiguë hypertriglycéridémique chez un enfant.

Acute pancreatitis in children is rare and is characterized by clinical, biological, and etiological polymorphism. We report the case of a 6-year-old child who underwent surgery for intestinal obstruction. The diagnosis of acute pancreatitis was made after the surgical exploration. Initial laboratory tests showed hyperlipasemia and mild hypertriglyceridemia. After 2 weeks of hospitalization, we found severe hypertriglyceridemia, which was considered the cause of acute pancreatitis. Based on a review of the literature, we emphasize the importance of considering acute pancreatitis in the diagnosis of abdominal pain in children. Hypertriglyceridemia is one of its rare causes and must be detected because specific treatments are needed. Blood triglyceride concentration in the initial phase can be misleading and the assay should be repeated after the acute episode.

Mesenteric appendicitis strangulating small bowel: an exceptional internal herniation.

Primitive internal hernias are a rare cause of intestinal obstruction. They are often paraduodenal even transmesocolic, but only rarely transomental. We present a rare case of an internal abdominal hernia in a young man. The small bowel was strangulated by an intra mesenteric appendicitis. This hernia was revealed by abdominal pain, nausea and vomiting. Plain X-ray of the abdomen showed dilated jejunal and ileal loops with multiple air-fluid levels. The diagnosis of appendicitis was suggested by ultrasound but the internal hernia was found only upon surgical exploration. An appendicectomy and adhesiolysis were performed. The patient recovered fully after 3 days, and had an uneventful postoperative course. The authors discuss the possible cause of this rare intestinal obstruction.

Primary pelvic hydatid cyst an unusual cause of cystic adnexal image (mass).

Hydatid cyst is a parasitic human infection which is endemic in North Africa. It is more frequently localized in the liver and the lung. Involvement of others sites is usually secondary to these primary localizations. We report 2 exceptional cases of primary pelvic hydatid cyst diagnosed respectively in a 37-year-old and in a 48-year-old women. These cases will focus on the different characteristics of the infection, and the benefit of including epidemiologic arguments in using the diagnostical approach of adnexal masses.

[Intracranial hydatid cysts in children: a report of 9 cases]. / Kystes hydatiques cérébraux de l'enfant (à propos de 9 cas).

PURPOSE: To illustrate the value of cross-sectional imaging (CT, MRI) for the diagnosis and follow-up of intracranial hydatid cysts in children. MATERIALS AND METHODS: Retrospective study of 9 cases of intracranial hydatid cysts in children seen over a period of 8 years. Precontrast and postcontrast 5 mm thick axial CT images were obtained in 7 cases. Noncontrast sagittal, axial and coronal T1W and T2W images were obtained in 2 cases. RESULTS: Mean patient age was 7.5 years. Intracranial hypertension was the main presenting clinical symptom. A single supratentorial cyst with significant mass effect upon the ventricular system and midline structures was observed in all cases. All patients underwent surgery with good outcome in all cases. CONCLUSION: CT is the imaging modality of choice for diagnosis and postoperative follow-up of intracranial hydatid cysts in children. MRI is most helpful for further characterization when multiple or atypical cysts are present to optimize management.

[Acute myocardial infarction revealing a pheochromocytoma: a case report]. / Infarctus du myocarde révélant un phéochromocytome : à propos d'un cas.

We report a case of a 38-year-old woman with pheochromocytoma admitted to our department for an acute myocardial infarction. The diagnosis of pheochromocytoma was evoked in view of the major pressure variations that appeared secondarily. Pheochromocytoma is mainly medullosurrenal tumour with clinical polymorphism; it can lead to severe cardiovascular disorders. Nevertheless, cardiac involvement is rarely in the foreground. Our clinical case illustrates the importance to evoke the diagnosis of pheochromocytoma in front of atypical manifestations of acute coronary syndrome so as to reduce mortality.

[Benign sacrococcygeal teratoma in a child: a case report with a review of the literature]. / Tératome sacrococcygien bénin chez l'enfant : revue de littérature à propos d'un cas.

Sacrococcygeal teratomas are rare congenital tumors, generally discovered at birth. These tumors are seldom observed in children. Radical resection must be performed to avoid potentially malignant recurrence even if the primary lesion is benign. We report a case in an 8-year-old girl who did not have a past medical history. Since the age of 2 years, she presented a progressive sacral tumefaction with no neurological deficit. The MRI showed a large sacrococcygeal cyst in hypointense-signal T1-weighted imaging with no contrast enhancement, and a hyperintense signal in T2-weighted imaging. At surgery, the tumor was totally removed. The intraoperative aspect was that of a viscous cyst. The histological study showed a sacrococcygeal teratoma.

[Giant cerebellar tuberculoma mimicking a malignant tumor]. / Tuberculome géant du cervelet simulant une tumeur maligne.

BACKGROUND: Isolated central nervous system (CNS) tuberculoma is rare. Central nervous system tuberculosis (TB) is associated with high morbidity and mortality despite modern methods of detection and treatment. The authors report a case of a giant cerebellar tuberculoma mimicking a malignant tumor and review the literature. OBSERVATION: A six-year-old girl, with no past medical history, vaccinated for her age, presented with a three-month history of occipitocervical cephalalgia, complicated by gait disturbances. The MRI showed a left cerebellar tumor suggestive of a medulloblastoma. At surgery, a nodular, avascular lesion was found and pathological examination confirmed tuberculoma. Intracranial tuberculoma is an uncommon variety of central nervous system tuberculosis. The prognosis is related to the rapidity of diagnosis, surgical resection and the complementary antituberculosis treatment. CONCLUSION: Intracranial tuberculoma is an uncommon variety of central nervous system infection. Prognosis is improved by a quick diagnosis, surgical removal, and associated antituberculoma therapy.

Molecular epidemiology of genital human papillomavirus and Chlamydia trachomatis among patients attending a genitourinary medicine clinic - will vaccines protect?

High-risk subtypes of human papillomavirus (HPV) are the main causative agents of cervical cancer, for which Chlamydia trachomatis (CT) may sometimes be a co-factor. Vaccines have been developed against some subtypes of human papillomavirus and a vaccine against CT is in development. The objective of this study was to determine the prevalence of the subtypes of HPV and CT in genitourinary (GU) medicine clinic attenders. In total, 1000 consecutive patients attending the GU clinic participated in this anonymized point-prevalence study. Urethral swabs from 437 men and urethral plus cervical swabs as a single specimen from 563 women were tested for the subtypes of both organisms. Nested major outer membrane protein (MOMP) polymerase chain reaction detected CT chromosomal DNA in 44/437 (10%) of the men and 73/563 (13%) of the women. Genotypes E, F, and D were the most common. In all, 55/437 (13%) of men and 244/563 (43%) of women were infected with at least one high-risk HPV type. In conclusion, the new HPV vaccines, Gardasil and Cervarix, would have protected against 58% and 45%, respectively, of the high-risk subtypes found in women in this population. The rate of high-risk HPV infection (43%) found in women in this study raises concern.

An internally controlled, one-step, real-time RT-PCR assay for norovirus detection and genogrouping.

BACKGROUND: Reverse transcription (RT)-PCR for norovirus detection is prone to false-negative results due to inhibitory substances in faeces. An internal control is needed to monitor extraction efficiency and to detect inhibition. OBJECTIVES: To further develop a one-step RT-PCR assay for norovirus detection/genogrouping by addition of MS2 bacteriophage as an internal control. STUDY DESIGN: Our norovirus RT-PCR assay was modified by addition of MS2 phage to the extraction tray and primers/probe for MS2 detection to the reaction mix. The effect of addition of MS2 phage and MS2 primers/probe on the sensitivity/specificity of the PCR assay was examined. RESULTS: The addition of MS2 as an internal control showed no loss of sensitivity or specificity for norovirus detection. CONCLUSIONS: A triplex, one-step, type-specific, real-time RT-PCR with MS2 internal control has been developed for use in routine laboratory diagnosis of norovirus infection.

The superiority of polymerase chain reaction over an amplified enzyme immunoassay for the detection of genital chlamydial infections.

BACKGROUND/OBJECTIVES: The polymer conjugate enhanced enzyme immunoassay (IDEIA) and Cobas Amplicor polymerase chain reaction Chlamydia trachomatis (CT) (Amplicor PCR) are two commonly used assays for the diagnosis of CT infection. The performance of these assays was compared for the diagnosis of genital CT infection among 1000 consecutive patients attending a genitourinary medicine (GUM) clinic. Confirmation of positive results and the clinical significance of the absence of cryptic plasmid in chlamydia on the diagnosis of infection by Amplicor PCR were also investigated. METHODS: IDEIA, Amplicor PCR, and two nested in-house PCR assays targeting cryptic plasmid and omp1 gene were performed on all samples. DNA from Amplicor PCR negative samples was pooled for in-house PCR assays. Each pool contained DNA from seven Amplicor PCR negative samples. RESULTS: Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and efficiency of IDEIA in the diagnosis of genital CT infection were 80%, 97%, 80%, 97%, and 95%, respectively. Sensitivity, specificity, PPV, NPV and efficiency of Amplicor PCR were 99%, 98%, 89%, 100%, and 98%, respectively. 16 (11%) of 144 Amplicor PCR positive results were identified as false positive by in-house PCR assays. No isolate of plasmid free CT was detected among the study population. CONCLUSIONS: IDEIA should not be used for the diagnosis of CT infection because of its poor sensitivity. Although the analytic specificity of Amplicor PCR was 98%, because of the adverse medical, social, and psychological impact of false positive results for patients, confirmation of Amplicor PCR positive results by a different assay with comparable sensitivity is essential. Amplification assays targeting cryptic plasmid are appropriate for the diagnosis of genital CT infections.

Mumps epididymo-orchitis with prolonged detection of virus in semen and the development of anti-sperm antibodies.

Epididymo-orchitis is the most common complication of mumps in post-pubertal men. A case of MMR vaccine failure, in whom mumps and mumps-associated unilateral epididymo-orchitis developed, is presented in this article. Mumps virus was isolated from the semen 14 days after onset and mumps RNA was detected in semen for up to 40 days using RT-PCR. Epididymo-orchitis was associated with transient but significant reduction in sperm count and severe abnormalities in sperm morphology. It also led to the appearance of anti-sperm antibodies, which may have potential long-term adverse effects on the patient's fertility. Sequencing of the SH gene of the virus showed this to be a new mumps genotype distinct from the virus circulating currently in the UK.

Detection of bacteraemia in patients with fever and neutropenia using 16S rRNA gene amplification by polymerase chain reaction.

Episodes of fever and neutropenia are common complications of treatment for cancer. The use of prophylactic and early empirical antibiotics has reduced mortality but decreases the sensitivity of diagnostic tests based on culture. The aim of this study was to determine the potential of a broad diagnostic approach (eubacterial) based on 16S rRNA gene amplification and sequencing to augment cultural methods of diagnosis of bacteraemia in patients with fever and neutropenia in a regional paediatric oncology centre. One hundred eleven patient-episodes of fever and neutropenia were evaluated during the study period, 17 of which were associated with positive blood cultures, as follows: Staphylococcus epidermidis (n = 6 episodes), Enterococcus faecium (n = 2), Streptococcus sanguis (n = 3), Streptococcus mitis (n = 3), Staphylococcus aureus (n = 1), Micrococcus spp. (n = 1), and Stenotrophomonas maltophilia (n = 1). Eubacterial polymerase chain reaction (PCR) detected bacterial DNA in nine of 11 blood culture-positive episodes for which a sample was available for PCR; the species identified by sequence analysis were identical to those derived from the conventional identification of the cultured isolates. Bacterial DNA was detected in 20 episodes (21 bacterial sequences) associated with negative blood cultures, 18 of which occurred in patients who were receiving antibiotics at the time of sample collection. The species presumptively identified by partial 16S rRNA gene sequencing were as follows: Pseudomonas spp. (n = 6 episodes), Acinetobacter spp. (n =5 ); Escherichia spp. (n = 3); Moraxella spp. (n = 3); Staphylococcus spp. (n = 2); Neisseria spp. (n = 1); and Bacillus spp. (n = 1). The results of this study suggest that molecular techniques can augment cultural methods in the diagnosis of bacteraemia in patients who have been treated with antibiotics.

Eubacterial approach to the diagnosis of bacterial infection.

Computed tomography revealed a frontal empyema in an 8 month old boy who presented with a prolonged convulsion. Before admission he had received oral antibiotics and all specimens were negative on culture. The presence of bacteria in cerebrospinal and empyema fluid was demonstrated using the polymerase chain reaction (PCR) with 16S rRNA gene primers. A presumptive identification of Streptococcus pneumoniae was made by comparison of PCR products with those derived from known bacteria using denaturing gradient gel electrophoresis.

Determination of penicillin susceptibility of Streptococcus pneumoniae using the polymerase chain reaction.

AIM: To develop a polymerase chain reaction (PCR) based method to detect penicillin susceptibility in isolates of Streptococcus pneumoniae (SP). METHOD: PCR primers were designed to amplify differential nucleotide sequences of the penicillin-binding protein (PBP) genes 2b, 2x, and 1a in penicillin susceptible and resistant strains of SP. Primers derived from the PBP 2x and 2b genes were designed to amplify products from penicillin susceptible S pneumoniae (PSSP), whereas primers derived from the PBP 1a gene were designed to amplify gene sequences of penicillin resistant S pneumoniae (PRSP). RESULTS: Two hundred and thirty clinical isolates of SP from the USA, UK, Kenya, Romania, and the Kingdom of Saudi Arabia were tested. Of the isolates, 116 were penicillin susceptible, 65 were intermediately resistant, and 49 were highly resistant. PCR identified 108 (93%) of 116 of PSSP isolates, 55 (85%) of 65 intermediately resistant isolates, and all of the 49 highly resistant isolates of SP. The susceptibility of 16 (7%) isolates could not be determined using PCR. All of these 16 isolates had a minimum inhibitory concentration (MIC) of penicillin < 1 mg/l. None of the highly resistant isolates was identified as penicillin susceptible by PCR, although two of the isolates with intermediate resistance (MIC = 0.125 mg/l) were. CONCLUSION: Using primers that differentially identify the genotypes of susceptible and resistant strains of SP, PCR provides a rapid method for determining the penicillin susceptibility of SP isolates and could potentially be used for testing clinical samples.

Changes in Bcl-2 and p53 expression in recurrent B-cell lymphomas.

The aim of this study was to investigate the changes involved in the evolution of nine cases of recurrent B-cell lymphomas. Using the polymerase chain reaction (PCR) on formalin-fixed, paraffin-embedded tissue from both the primary and the recurrent lymphoma of each case, monoclonality was demonstrated in every tumour. In all nine cases, the recurrent lymphoma was shown to belong to the same clone as the primary lymphoma. Eight of these cases were then investigated by immunohistochemistry for changes in Bcl-2 and p53 expression. Five out of eight of the primary lymphomas showed Bcl-2 overexpression. Two of the three cases initially negative for Bcl-2 expression became positive in the recurrence. One out of eight of the primary lymphomas was positive for p53 expression. Of the seven negative cases, one became positive for p53 expression in the recurrence. Both of the p53-positive cases showed high-grade histology. This study shows that Bcl-2 overexpression is probably an important early event in the development of B-cell lymphomas, although it may occur as a post-neoplastic event. p53 mutation is probably more important as a late event and may be associated with high-grade transformation.

Application of 16S rRNA gene PCR to study bowel flora of preterm infants with and without necrotizing enterocolitis.

The purpose of the present study was to determine the extent to which bacteria not detected by culture contribute to the microbial flora of the bowel of preterm infants with and without neonatal necrotizing enterocolitis (NEC). Fecal samples from 32 preterm infants in special care baby units including samples from 10 infants with NEC were examined by culture and PCR amplification of the 16S rRNA gene (rDNA). The 16S rDNA V3 region was amplified with eubacterial primers, and the amplification products derived from the fecal sample DNA were compared with the products from individual cultured isolates by PCR and denaturing gradient gel electrophoresis (PCR-DGGE), allowing the DNA from uncultured bacteria to be identified. For the 22 infants without NEC weekly samples were examined for a mean of 5.3 postnatal weeks. The total number of types detected by culture combined with PCR-DGGE was 10.1 per infant, of which PCR-DGGE contributed 10.4% of the types identified. Additional types detected by PCR-DGGE were found in 14 (63.6%) of the 22 infants. The majority of the sequences associated with uncultured bacteria showed > 90% 16S rDNA sequence identity with sequences from culturable human enteric flora, and all were found in single infants with the exception of sequences indistinguishable by DGGE from seven infants. These sequences showed > 90% sequence identity with the 16S rDNA of Streptococcus salivarius and may have been derived from upper gastrointestinal or respiratory tract flora. In the present study uncultured bacteria detected by PCR-DGGE were no more frequent in fecal samples from infants with NEC than in samples from infants without NEC, although these findings do not exclude the possibility of unrecognized bacteria associated with the mucosa of the small intestine of infants with NEC.