Application of a recombinase polymerase amplification (RPA) assay and pilot field testing for Giardia duodenalis at Lake Albert, Uganda.

BACKGROUND: Giardia duodenalis is a gastrointestinal protozoan causing 184 million cases of giardiasis worldwide annually. Detection is by microscopy or coproantigen assays, although sensitivity is often compromised by intermittent shedding of cysts or trophozoites, or operator expertise. Therefore, for enhanced surveillance field-applicable, point-of-care (POC), molecular assays are needed. Our aims were to: (i) optimise the recombinase polymerase amplification (RPA) assay for the isothermal amplification of the G. duodenalis ß-giardin gene from trophozoites and cysts, using published primer and probes; and (ii) perform a pilot field validation of RPA at a field station in a resource-poor setting, on DNA extracted from stool samples from schoolchildren in villages around Lake Albert, Uganda. Results were compared to an established laboratory small subunit ribosomal RNA (SSU rDNA) qPCR assay with additional testing using a qPCR targeting the triose phosphate isomerase (tpi) DNA regions that can distinguish G. duodenalis of two different assemblages (A and B), which are human-specific. RESULTS: Initial optimisation resulted in the successful amplification of predicted RPA products from G. duodenalis-purified gDNA, producing a double-labelled amplicon detected using lateral flow strips. In the field setting, of 129 stool samples, 49 (37.9%) were positive using the Giardia/Cryptosporidium QuikChek coproantigen test; however, the RPA assay when conducted in the field was positive for a single stool sample. Subsequent molecular screening in the laboratory on a subset (n = 73) of the samples demonstrated better results with 21 (28.8%) RPA positive. The SSU rDNA qPCR assay resulted in 30/129 (23.3%) positive samples; 18 out of 73 (24.7%) were assemblage typed (9 assemblage A; 5 assemblage B; and 4 mixed A+B). Compared with the SSU rDNA qPCR, QuikChek was more sensitive than RPA (85.7 vs 61.9%), but with similar specificities (80.8 vs 84.6%). In comparison to QuikChek, RPA had 46.4% sensitivity and 82.2% specificity. CONCLUSIONS: To the best of our knowledge, this is the first in-field and comparative laboratory validation of RPA for giardiasis in low resource settings. Further refinement and technology transfer, specifically in relation to stool sample preparation, will be needed to implement this assay in the field, which could assist better detection of asymptomatic Giardia infections.

Molecular detection of equine trypanosomiasis in the Riyadh Province of Saudi Arabia.

We conducted a cross-sectional study to detect trypanosome infections of horses and donkeys in the Riyadh Province of Saudi Arabia. DNA was extracted from blood samples collected from 368 horses and 142 donkeys, and subjected to universal first ribosomal internal transcribed spacer region (ITS1)-PCR followed by Trypanosoma evansi species-specific RoTat1.2-PCR. The universal ITS1-PCR revealed T. evansi infection in horses ( n = 12; 3.3%) and donkeys ( n = 4; 2.8%). There was no significant effect of sex or age on the prevalence of trypanosomiasis in horses or donkeys. Application of the RoTat1.2-PCR revealed that the RoTat1.2 VSG gene was absent from the positive ITS1-PCR samples of 3 horses and 1 donkey. This discrepancy could be explained by the circulation of T. evansi type B in Saudi Arabia; however, this suspicion requires confirmation.

Comparison of the clinical effectiveness of thermal cure and rehabilitation in knee osteoarthritis. A randomized therapeutic trial.

OBJECTIVE: To compare the benefits of a thermal cure and non-thermal rehabilitation in treatment of knee osteoarthritis (KOA). METHODS: Randomized therapeutic trial including patients with knee osteoarthritis (American College of Rheumatology criteria). Patients were randomly divided into two groups. Spa treatment consisted of underwater shower, massage-jet showers, hydromassage, pool rehabilitation and peloid therapy. Non-thermal rehabilitation consisted of analgesic physiotherapy, muscle strengthening and group physical rehabilitation. A blinded evaluation was carried out at day 21 and 12months following treatment. It was based on the visual analogic scale of pain (VAS), which represented the primary endpoint. RESULTS: Two hundred and forty patients were included (February-June 2005). The spa treatment and non-thermal rehabilitation groups included 119 and 121 patients respectively. Two hundred and thirty-three patients completed their treatments. Significant improvement of the visual analogic scale of pain was noted in the thermal cure group (61.6±15 at day 0 versus 46.5±22.4 at 12months, P<0.001), but not in the non-thermal group (64.1±15 at day 0 versus 62±29 at 12months, P=0.68). At day 21, comparison of the two groups revealed no significant difference on the VAS (P=0.08). However, at 12months, the thermal cure group was significantly more improved (P=0.000). CONCLUSION: In our study, crenobalneotherapy had resulted, at 12months, in more pronounced long-term improvement of the painful symptoms of KOA than had non-thermal rehabilitation.

[Intraosseous capillary hemangioma of the radial bone]. / Une tumeur osseuse rare au radius : hémangiome capillaire.

Hemangioma is a rare benign vascular tumor composed of dystrophic capillaries, preferentially located in the spinal region (70 %). Bony localizations are exceptional, less than 1 % of cases in large series of bone tumors. We report the case of a 35-year-old male who complained of pain the right forearm for two months. The pain was of an inflammatory type and was triggered by palpation and mobilization. The plain x-ray revealed a bony defect involving the posterior cortical of the mid-third of the radius. Magnetic resonance imaging revealed a defect of the shaft cortex without involvement of the central part of the bone, the endostium or the soft tissue. Surgical resection was limited to the tumor. Histology reported intraosseous capillary hemangioma. The course was favorable with bone healing and gap filling, with normal elbow and wrist function. The patient resumed occupational activities three months after surgery. This case illustrates a rare localization of this tumor rarely reported in major series.

[Juvenile idiopathic arthritis treated by methotrexate: result of treatment and tolerance]. / Les arthrites juvéniles idiopathiques traitées par méthotrexate. réponse au traitement et tolerance à moyen terme.

The MTX has a place of choice in the treatment of the AJI with good observance and the absence of grave side effects. In a retrospective study, 14 cases of AJI having benefited from a treatment by MTX were included in the study. Answer to the treatment was estimated at the 6-th month on the criteria of RINTO. The group included in the therapeutic evaluation, is formed by 10 boys and by 4 girls. The average age of the beginning of the disease is of 6 years (2 - 11). The shape of the beginning is systematic in 7 cases and polyarticular in 5 cases and oligoarticular in 2 cases.. The average duration of the treatment by MTX is of 36 months (9 - 66). Answer to the MTX in the 6-th month was noted in 7 cases, under a MTX's average dose of 7, 14mg / m2 / week. Unwanted effects were observed in 7 cases and have not in any case led to the stop of the treatment

Myositis ossificans circumscripta of the knee improved by alendronate.

Myositis ossificans circumscripta (MOC) is a rare benign disorder characterized by heterotopic ossification of soft tissues. We report a case in a 21-year-old male who presented with knee pain after strenuous physical activity. Evidence of inflammation of the knee and a hard swelling in the subquadricipital area were the main physical findings. The initial plain radiographs showed a subpatellar soft tissue mass containing flocculent calcifications. Over the next 2 months, the calcific deposits organized into a calcified mass. Magnetic resonance imaging findings were nonspecific, with a mass within the vastus medialis muscle generating low signal on T1 images and high signal on T2 images, without postgadolinium enhancement. Computed tomography showed several calcific deposits arranged in a ring, strongly suggesting MOC. Histological examination of a biopsy specimen confirmed this diagnosis. Alendronate therapy given for 6 months was followed by marked clinical improvement, obviating the need for surgery.

[Osteoid osteoma of the tarsal navicular bone: an uncommon localization]. / Ostéome ostéoïde de l'os naviculaire tarsien: une localisation rare.

We report a case of subchondral osteoid osteoma of the tarsal navicular bone. Osteoid osteomas are often located in the foot, mainly in the talus. This is the first report in the literature of a tarsal navicular bone localization. Diagnosis was difficult and established late due to misleading clinical presentation simulating mediotarsal arthritis.

Autoantibodies to ribosomal P proteins in systemic lupus erythematosus.

UNLABELLED: Autoantibodies to ribosomal P proteins (anti-P) are directed against three ribosomal phosphoproteins (P0, P1, and P2) and are believed to be specific markers for systemic lupus erythematosus (SLE), particularly with neuropsychiatric manifestations. The objective of this study was to determine the rate of occurrence and clinical significance of anti-P in Tunisian patients. METHODS: We included 100 Tunisian patients with SLE whose sera were received between January 1998 and December 2000 by the immunology laboratory of the Charles Nicolle Teaching Hospital in Tunis, Tunisia, and were tested for anti-P. An immunodot technique based on a synthetic protein shared by all ribosomal phosphoproteins was used. RESULTS: Sera from 22 (22%) patients were positive for anti-P. This rate is consistent with the literature. We found no statistically significant associations between anti-P and specific manifestations of SLE. CONCLUSION. We found that anti-P were significantly associated with anti-Sm and anti-RNP, as reported previously. The significance of these associations remains debatable.

[Rheumatic spondylodiscitis in spondylarthropathies]. / Spondylodiscite rhumatismale au cours des spondylarthropathies.

The involvement of enthesis like disc space is the typical lesion of spondylarthropathies. Out of 240 patients with spondylarthropathies, 10 (7 M, 3 W; mean age: 37 years) have spondylodiscitis, affecting a total of 13 disc spaces: 1 cervical, 2 thoracic, 10 lumbar. The mean duration of spondylarthropathy at the time of diagnosis of spondylodiscitis is 10 years. Spondylodiscitis has a variable clinical presentation: painful (6 cases), asymptomatic (4 cases). Its coexistence with typical radiographic signs of spondylarthropathy added to the exclusion of bacterial origin, permit to link it to spondylarthropathy. Its evolution is generally favourable.

[Anti-ribosomal antibodies during hepatitis C]. / Anticorps anti-ribosomes au cours d'une hépatite C.

We report the case of a young Tunisian woman hospitalised for reasons of general ill health. A immunological check-up revealed autoimmune disturbances and in particular anti-ribosomal P antibodies. We discuss the possible relation of these antibodies to the hepatitis C infection from which the patient was also suffering.

[Hereditary chondrocalcinosis in Tunisia. Apropos of 3 families]. / Chondrocalcinose héréditaire en Tunisie. A propos de trois familles.

Hereditary chondrocalcinosis was reported in three Tunisian families. The clinical manifestations appeared early in life and extensive radiologic involvement was apparent. The mode of inheritance seemed to be autosomal with incomplete penetrance. In one family HLA typing revealed that all affected subjects had the haplotype A1 B12 DR3.