Matthew R. Watts

Real-Time Polymerase Chain Reaction for Detection of Strongyloides stercoralis in Stool

The use of real-time polymerase chain reaction (PCR) for detection of Strongyloides stercoralis in stool has recently been described. We compared five DNA extraction methods by using normal human stool spiked with Strongyloides ratti and tested by using a real-time PCR. The PowerSoil kit was found to be the best technique in terms of sensitivity and ease of use. The PCR detected DNA extracted from one spiked S. ratti larva diluted 10⁻². The PowerSoil kit was then used to extract DNA from 160 human survey samples. All culture positive specimens with a high and moderate larval load were identified by real-time PCR, but only 15% of specimens with low larval load were positive. Specificity was greater than 99%. The combination of the PowerSoil kit and real-time PCR reliably detected high to moderate larval numbers of S. stercoralis in stools but was less sensitive when the larval load was low.

A Loop-Mediated Isothermal Amplification (LAMP) Assay for Strongyloides stercoralis in Stool That Uses a Visual Detection Method with SYTO-82 Fluorescent Dye

An assay to detect Strongyloides stercoralis in stool specimens was developed using the loop-mediated isothermal amplification (LAMP) method. Primers were based on the 28S ribosomal subunit gene. The reaction conditions were optimized and SYTO-82 fluorescent dye was used to allow real-time and visual detection of the product. The product identity was confirmed with restriction enzyme digestion, cloning, and sequence analysis. The assay was specific when tested against DNA from bacteria, fungi and parasites, and 30 normal stool samples. Analytical sensitivity was to < 10 copies of target sequence in a plasmid and up to a 10(-2) dilution of DNA extracted from a Strongyloides ratti larva spiked into stool. Sensitivity was increased when further dilutions were made in water, indicative of reduced reaction inhibition. Twenty-seven of 28 stool samples microscopy and polymerase chain reaction positive for S. stercoralis were positive with the LAMP method. On the basis of these findings, the assay warrants further clinical validation.

Anncaliia algerae Microsporidial Myositis

Immunosuppression is a risk factor for serious infection in humans.

Nocardia infections of the face and neck.

Involvement of the soft tissues of the face and neck by Nocardia spp. is uncommon. We review the epidemiology, clinical features, diagnosis, and management of such infections in the setting of primary cutaneous nocardiosis and disseminated disease. Although immune compromise is an important risk factor for these infections, they also occur in healthy individuals. Infection may arise through direct inoculation following injury or by hematogenous spread from a primary site, usually the lung. The rare variant of lymphocutaneous disease—cervicofacial nocardiosis—typically affects children, but can occur in adults. The diagnosis is made by conventional microscopy and culture, but radiological imaging is useful to delineate disease extent, and molecular methods are increasingly assisting the diagnosis by providing rapid detection and identification of the pathogen. Sulfonamides remain the preferred treatment for many cases and are an important component of the therapeutic armamentarium. Other therapeutic options include minocycline, the carbapenems, and linezolid.

Treatment of pseudoepitheliomatous, keratotic, and micaceous balanitis with topical photodynamic therapy.

Pseudoepitheliomatous, keratotic, and micaceous balanitis (PKMB) is an extremely rare condition occurring over the glans in older men who undergo circumcision late in life. Its exact etiology is unknown.

Coccidioidal septic arthritis: lessons learned from a clinical and laboratory perspective.

The Medical Journal of Australia ISSN: 0025729X 18 June 2012 196 11 705-706 ©The Medical Journal of Australia 2012 www.mja.com.au Case reports — lessons from Practice MJA 2012; 196: 705–706 doi: 10.5694/mja11.10533 Coccidioidomycosis (“pseudotu sive fungal infection caused by immitis and C. posadasii). It is acquired in Australia, being end sph re deserts, primarily in the so States, and Mexico. After respirat Imported pathogens are increasingly recognised as potential biosecurity concerns, but may be overlooked when the clinical syndrome and causative organism are unexpected. This case highlights the need to consider an infectious aetiology in an older migrant with a chronic inflammatory monoarthritis. It emphasises the need for good communication between laboratory disciplines, and demonstrates preliminary success with a combined surgical–medical– pathology–pharmacological approach. berculosis”) is an invaCoccidioides species (C. not autochthonously emic to western-hemiuth-west of the United ory inoculation, arthroconidia (hyphal forms) transform into multinucleated spherules, which rupture, propagating new spherules.5 Crucial diagnostic information obtained through the histopathological detection of spherules in this patient (Box 2) was rapidly relayed to the microbiology laboratory. Coccidioides spp are classified as Risk Group 3 pathogens in Australia (as is Bacillus anthracis).6 Analytical work performed on these pathogens must be carried out in a biological safety cabinet in a physical containment level 3 sealed laboratory with negative air pressure.6 As one of the ten most common laboratory-acquired infections,7 with laboratory attack rates exceeding those of natural exposure,8 travel history to an endemic area is crucial information for laboratory staff.3,9 Suspected C. immitis colonies take up to 5 days to manifest with “typical” morphological Clinical record An 81-year-old non-English-speaking Filipino woman with type 2 diabetes mellitus presented with left knee pain and swelling of 18 months’ duration. Her mobility was restricted to less than 2m because of pain. Synovial fluid obtained by needle aspiration during a recent visit to the Philippines had yielded a white cell count of 12200 cells/L (55% polymorphonucleocytes; reference interval [RI] not available) and no bacterial organisms. On her return to Australia, she was referred to an orthopaedic surgeon. Physical examination revealed chronically discharging wounds at previous aspiration sites of the medial and lateral aspects of the knee, which was swollen and tender but with good range of movement. The patient did not have a fever, and her C-reactive protein level was 18 mg/L (RI, < 10 mg/L)]. Magnetic resonance imaging demonstrated florid erosive synovitis, considered most consistent with rheumatoid or seronegative arthritis, although bone oedema was noted, suggesting osteomyelitis (Box 1). Culture of material from the sinuses for bacteria was negative.

A 12-Year Retrospective Study of Invasive Amoebiasis in Western Sydney: Evidence of Local Acquisition

In Australia, amoebiasis is thought to occur in travellers, immigrants from endemic areas, and among men who have sex with men. Prevalence of amoebiasis in communities with immigrants from Entamoeba histolytica-endemic countries is unknown. The present study is a retrospective case series analysis of patients with laboratory-confirmed amoebiasis from Western Sydney Local Health District, Australia, between years 2005 and 2016. Forty-nine patients with amoebiasis were identified, resulting in an estimated annual incidence of up to 1.1 cases per 100,000 adults. Many were born in Australia (15/47) and India (12/47). Three patients (3/37) had no history of overseas travel, two others had not travelled to an endemic country, and an additional two had a very remote history of overseas travel; one died of fulminant amoebic colitis. Three patients (3/16) were employed in the food industry and one had a history of colonic irrigation in an Australian ‘wellness clinic’. Patients had invasive amoebiasis with either liver abscess (41/48) or colitis (7/48), diagnosed most commonly by serology. Invasive procedures were common, including aspiration of liver abscess (28/41), colonoscopy (11/49), and partial hepatectomy (1/49). Although rare, local acquisition of amoebiasis occurs in Western Sydney and contributes to significant morbidity and hospital admissions.

Anncaliia algerae Microsporidial Myositis, New South Wales, Australia

We describe the successful management of Anncaliia algerae microsporidial myositis in a man with graft versus host disease after hemopoietic stem cell transplantation. We also summarize clinical presentation and management approaches and discuss the importance of research into the acquisition of this infection and strategies for prevention.

Cutaneous non-tuberculous mycobacteria in Western Sydney, Australia. Population study 1996-2013

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Extensive cystic hydatid disease.

SnapShot A 46-year-old woman, who had lived in rural Australia as a child, presented with abdominal distension, jaundice and cachexia. Abdominal computed tomography showed extensive cystic disease of the liver, peritoneum, ovaries and uterus; well circumscribed coarsely calcified cysts were visible in the abdomen and pelvis (Figure A). Results of hydatid serological testing were consistent with Echinococcus granulosus infection (indirect haemagglutination titre, 256). The patient was managed with preoperative albendazole therapy, extensive surgical excision, hypertonic saline irrigation, and decompression of the ruptured left hepatic duct with a T-tube. Microscopy of cystic fluid aspirated during surgery showed viable evaginating protoscolices (Figure B; original magnification,  400). Longterm suppressive therapy — albendazole 400 mg twice daily plus praziquantel 40 mg/kg twice weekly — was initiated with good effect.