Sensitivity and specifi city of HAT Sero-K-SeT, a rapid diagnostic test for serodiagnosis of sleeping sickness caused by Trypanosoma brucei gambiense: a case-control study

Background Human African trypanosomiasis (HAT) is a life-threatening infection aff ecting rural populations in sub- Saharan Africa. Large-scale population screening by antibody detection with the Card Agglutination Test for Trypanosomiasis (CATT)/Trypanosoma brucei (T b) gambiense helped reduce the number of reported cases of gambiense HAT to fewer than 10 000 in 2011. Because low case numbers lead to decreased cost-eff ectiveness of such active screening, we aimed to assess diagnostic accuracy of a rapid serodiagnostic test (HAT Sero-K-SeT) applicable in primary health-care centres. Methods In our case-control study, we assessed participants older than 11 years who presented for HAT Sero-K-SeT and CATT/T b gambiense at primary care centres or to mobile teams (and existing patients with confi rmed disease status at these centres) in Bandundu Province, DR Congo. We defi ned cases as patients with trypanosomes that had been identifi ed in lymph node aspirate, blood, or cerebrospinal fl uid. During screening, we recruited controls without previous history of HAT or detectable trypanosomes in blood or lymph who resided in the same area as the cases. We assessed diagnostic accuracy of three antibody detection tests for gambiense HAT: HAT Sero-K-SeT and CATT/T b gambiense (done with venous blood at the primary care centres) and immune trypanolysis (done with plasma at the Institute of Tropical Medicine, Antwerp, Belgium). Findings Between June 6, 2012, and Feb 25, 2013, we included 134 cases and 356 controls. HAT Sero-K-SeT had a sensitivity of 0·985 (132 true positives, 95% CI 0·947–0·996) and a specifi city of 0·986 (351 true negatives, 0·968–0·994), which did not diff er signifi cantly from CATT/T b gambiense (sensitivity 95% CI 0·955, 95% CI 0·906–0·979 [128 true positives] and specifi city 0·972, 0·949–0·985 [346 true negatives]) or immune trypanolysis (sensitivity 0·985, 0·947–0·996 [132 true positives] and specifi city 0·980, 0·960–0·990 [349 true negatives]). Interpretation The diagnostic accuracy of HAT Sero-K-SeT is adequate for T b gambiense antibody detection in local health centres and could be used for active screening whenever a cold chain and electricity supply are unavailable and CATT/T b gambiense cannot be done

Development and Potential Usefulness of the COVID-19 Ag Respi-Strip Diagnostic Assay in a Pandemic Context

Introduction: COVID-19 Ag Respi-Strip, an immunochromatographic (ICT) assay for the rapid detection of SARS-CoV-2 antigen on nasopharyngeal specimen, has been developed to identify positive COVID-19 patients allowing prompt clinical and quarantine decisions. In this original research article, we describe the conception, the analytical and clinical performances as well as the risk management of implementing the COVID-19 Ag Respi-Strip in a diagnostic decision algorithm. Materials and Methods: Development of the COVID-19 Ag Respi-Strip resulted in a ready-to-use ICT assay based on a membrane technology with colloidal gold nanoparticles using monoclonal antibodies directed against the SARS-CoV and SARS-CoV-2 highly conserved nucleoprotein antigen. Four hundred observations were recorded for the analytical performance study and thirty tests were analyzed for the crossreactivity study. The clinical performance study was performed in a retrospective multicentric evaluation on aliquots of 328 nasopharyngeal samples. COVID-19 Ag Respi-Strip results were compared with qRT-PCR as golden standard for COVID-19 diagnostics. Results: In the analytical performance study, the reproducibility showed a between-observer disagreement of 1.7%, a robustness of 98%, an overall satisfying user friendliness and no cross-reactivity with other virus-infected nasopharyngeal samples. In the clinical performance study performed in three different clinical laboratories during the ascendant phase of the epidemiological curve, we found an overall sensitivity and specificity of 57.6 and 99.5%, respectively with an accuracy of 82.6%. The cut-off of the ICT was found at CT < 22. User-friendliness analysis and risk management assessment through Ishikawa diagram demonstrate that COVID-19 Ag Respi-Strip may be implemented in clinical laboratories according to biosafety recommendations. Conclusion: The COVID-19 Ag Respi-Strip represents a promising rapid SARS-CoV-2 antigen assay for the first-line diagnosis of COVID-19 in 15min at the peak of the pandemic. Its role in the proposed diagnostic algorithm is complementary to the currently-used molecular techniques

T. cruzi OligoC-TesT: A Simplified and Standardized Polymerase Chain Reaction Format for Diagnosis of Chagas Disease

Chagas disease (American trypanosomiasis) is caused by the protozoan parasite Trypanosoma cruzi and represents a major public health problem in Latin America. Furthermore, growing human population movements extend the disease distribution to regions outside the South American continent. Accurate diagnosis is crucial in patient care and in preventing transmission through blood transfusion, organ transplantation, or vertical transmission from mother to child. Routine diagnosis of Trypanosoma cruzi infection generally is based on detection of the host's antibodies against the parasite. However, antibody detection tests are liable to specificity problems and are of limited use in assessing treatment outcome and congenital infections. The introduction of the polymerase chain reaction (PCR) to amplify specific DNA sequences opened promising diagnostic perspectives. Despite its reported high sensitivity and specificity, broad use of the PCR technique in diagnosis of Chagas disease is hampered by its complexity and the lack of any standardization. We here present the development and evaluation of the T. cruzi OligoC-TesT, a simple and standardized dipstick format for detection of PCR amplified T. cruzi DNA. The new tool is an important step towards simplified and standardized molecular diagnosis of Chagas disease

Survival, immune responses and tissue cyst production in outbred (Swiss white) and inbred (CBA/Ca) strains of mice experimentally infected with Neospora caninum tachyzoites

The present work compared inbred (CBA/Ca) and outbred (Swiss white) strains of mice for their capacity to cope with a Neospora caninum infection and to consistently produce tissue cysts. In each experiment Swiss white and CBA/Ca mice were given three different doses of NC-1 tachyzoites. Lymphoproliferative and Immoral responses as well as cytokine production were evaluated eight weeks after infection (PI) whereas tissue cyst production and histopathology were assessed 4, 6 and 10 weeks PI in immunosuppressed mice. Tissue cysts were observed 10 weeks after infection only in CBA/Ca mice receiving the two highest inoculum doses. Furthermore this strain showed the highest specific lymphoproliferative response. A mixed cytokine response with elevated IFN-gamma and fairly low IL-4 and IL-10 secretion was recorded. In both strains, no lesions were observed in the tissues of infected mice. This study indicates that CBA/Ca female mice infected with 5 x 10(6) NC-1 tachyzoites represent a useful model for the study of specific maternal immune responses in pregnant animals

In vitro tests for evaluation of the hatchability of the eggs of Psoroptes mites following exposure to acaricidal compounds.

Three in vitro assays for the evaluation of the hatchability of the eggs of the mite Psoroptes ovis (Hering) (Acari: Psoroptidae) are described. Preliminary trials showed that hatching occurs at very high rates when eggs are incubated under conditions of high humidity, on a liquid medium and in agarose dishes. These three protocols were compared, taking into account the ease of preparation, follow-up and accuracy of counting. The best protocol was found to be the use of agarose dishes. It was accurate, easy to carry out and reproducible for further evaluation of existing or potentially new compounds against both adults and eggs of Psoroptes spp. The acaricidal properties of phoxim and amitraz were then evaluated against eggs and adults using the three protocols. Results showed that for both drugs, in vitro adulticidal activity was complete, whereas the in vitro ovicidal activity was only partial. Nevertheless, efficacy of amitraz against both adults and eggs was shown to be higher than that of phoxim

Humoral Immune Response in Calves to Single-Dose, Trickle and Challenge Infections with Fasciola Hepatica

In cattle experimentally infected with Fasciola hepatica, parasite specific IgG1 and IgG2 responses were studied. Additionally parasite specific IgE production was assessed by the Passive Cutaneous Anaphylaxis reaction. The primary infection was administered either as a single-dose or as a trickle infection over a 4-week period. Animals were challenged 4 months later. Titres of IgG1 and IgG2 against excretory-secretory parasite products (FhESAg), and against a whole-worm extract (FhSomAg) were measured by enzyme-linked immunosorbent assay (ELISA) in relation to weight gain, serum hepatic enzyme levels, and fluke infection rate. At necropsy, the mean number of flukes recovered was similar in both infected groups. The two ELISAs specific for bovine IgG1 showed analogous sensitivity and specificity (92% and 94%). Cross-reactivity was observed towards Echinococcus granulosus, Cysticercus tenuicollis, and C. ovis but not towards C. bovis, Cooperia spp., and Ostertagia spp. FhESAg gave rise to apparently more stable specific IgG1 titres as compared to FhSomAg. Mean IgG1 titres were significantly higher in the single-dose-infected group than in the trickle-infected group during the early migratory phase of the infection (week 2 to week 4 (FhSomAg) or week 6 (FhESAg)). IgG2 values were consistently lower than IgG1 levels. The kinetic response of both isotypes yielded a similar pattern. Specific IgE antibodies were detected in cattle of both infected groups from week 2 post-primary infection (PPI) onwards. The mean serum glutamate dehydrogenase (GLDH) and gamma-glutamyl transferase (gammaGT) activities were significantly higher in the single-dose-infected group for 3 weeks around peak levels (12-14 weeks PPI and 14-16 weeks PPI for GLDH and gammaGT respectively). Western blotting revealed a major antigenic fraction in FhESAg (26-30 kDa) recognized specifically by sera from F. hepatica infected calves as early as 6-8 weeks PPI. Experimental challenge caused no statistically significant modification of any parameter (IgG1 and IgG2 titres, enzymatic activities, immunoblotting) used to monitor the course of the infection. No correlation was found between fluke size and number, and antibody titres, suggesting that IgG1 production has little protective effect against F. hepatica infection

Ultrastructural morphology of the male and female genital tracts of Psoroptes spp. (Acari : Astigmata : Psoroptidae)

The structure of the male and female genital systems of the astigmatid mite Psoroptes ovis (Hering) is described. The male genital system is composed of a paired testis, fused at its proximal part, two vasa deferentia, an ejaculatory duct, into which a single accessory gland opens, and a copulatory organ. The testis is characterized by a peripheric syncytial cell surrounding spermatogonia, spermatocytes, spermatids and spermatozoa which are distributed regularly in the gonad according to the sequence of spermatogenesis. The female genital system consists of a copulatory pore (the bursa copulatrix), a seminal receptacle, paired ovaries and oviducts, a glandular uterus and an ovipositor which leads to the oviporus. Ovaries are composed of somatic cells, germ cells and a central cell, with a multilobular nucleus, connected to oocytes by a stalk. Similarities with other astigmatic mites belonging to Psoroptidia and Acaridia are also discussed

Evaluation of two new commercial immunochromatographic assays for the rapid detection of OXA-48 and KPC carbapenemases from cultured bacteria.

BACKGROUND: Rapid detection and confirmation of carbapenemases remains very challenging for diagnostic laboratories. OBJECTIVES: The objective of this study was to assess the performance of two new immunochromatographic (IC) commercial assays for the rapid detection of OXA-48-producing and KPC-producing Enterobacteriaceae in pure bacterial isolates. METHODS: A panel of 92 bacterial isolates predominantly including carbapenem-non-susceptible Enterobacteriaceae with previously defined carbapenem resistance mechanisms was tested. Then, 342 consecutive carbapenem-non-susceptible Enterobacteriaceae isolates referred to the reference laboratory were investigated prospectively in parallel with other phenotypic tests and with multiplex PCR and sequencing as the gold standard. RESULTS: In the collection panel, each of the two IC assays correctly detected all 30 OXA-48-like-producing isolates and 25 KPC-producing isolates, whatever the species, their association with other β-lactamases and the level of resistance to carbapenems. All other carbapenemase producers and all non-carbapenemase-producing isolates yielded negative results with both tests. In the prospective evaluation, all OXA-48-like-producing Enterobacteriaceae isolates (n = 130) and KPC-producing Enterobacteriaceae isolates (n = 33) were correctly detected by the individual IC assays, while 179 non-OXA-48-like-producing and non-KPC-producing strains (137 non-carbapenemase producers and 42 isolates belonging to other carbapenemase family types) yielded negative results. Thus, each assay yielded 100% sensitivity and 100% specificity for the detection of OXA-48-like or KPC enzymes, respectively, at 15 min. CONCLUSIONS: The two IC assays allow rapid and reliable direct confirmation of OXA-48 and KPC carbapenemases from culture colonies and appear to be very useful additions to the existing tests, obviating the need for more costly characterization by molecular amplification methods

Use of a serum-free medium to produce in vitro Neospora caninum and Toxoplasma gondii tachyzoites on Vero cells

Neospora caninum and Toxoplasma gondii are cyst-forming coccidian parasites of human and veterinary clinical relevance. In vitro cultivation of the protozoans using Vero cells is usually performed in order to produce antigenic materials. Quantitative and qualitative comparisons of Vero cells grown in RPMI medium supplemented either with foetal calf serum (FCS), horse serum (HS) or a specific serum-free additive (DefCell) were performed. A serum-free cell culture system used to propagate N. caninum (NC-1 isolate) and T. gondii tachyzoites (Rh stain) were compared with the other two cell culture systems. FCS supplemented media was found to be more effective than the others in promoting Vero cells and N. caninum tachyzoites. However, it was found unable to support adequate T. gondii tachyzoite proliferation. Vero cells, T. gondii and N. caninum tachyzoite production gave similar growth patterns with either HS or DefCell supplemented media. Defcell was considered as a good alternative to supplement culture medium.Utilisation d'un milieu sans sérum pour la production in vitro de tachyzoites de Neospora caninum et Toxoplasma gondii sur cellules Vero. Neospora caninum et Toxoplasma gondii sont deux sporozoaires présentant un intérêt en médecine humaine et vétérinaire. La culture in vitro utilisant, entre autres, les cellules Vero comme support de la multiplication des deux parasites, est généralement employée en vue de l'obtention de tachyzoïtes. Au cours de cette étude, une évaluation quantitative et qualitative de la production de cellules Vero dans un milieu complémenté en sérum de veau foetal, de cheval ou bien avec un additif (DefCell) exempt de sérum, a été réalisée. Les trois types de milieu de culture ont également été employés et comparés dans le cadre de la production sur cellules Vero de tachyzoïtes de T. gondii (souche RH) et de N. caninum (isolat NC-1). Le milieu complémenté en sérum de veau fœtal s'est révélé être le plus adéquat pour la croissance de cellules Vero et la production de tachyzoïtes de N. caninum. Cependant, ce milieu s'est avéré incapable d'assurer une production optimale de tachyzoïtes de T. gondii. La production de cellules Vero, ainsi que de tachyzoïtes de T. gondii et de N. caninum, a présenté des caractéristiques communes en milieu complémenté en sérum de cheval et en DefCell. Ce dernier s'est révélé être une bonne alternative au sérum de veau foetal et de cheval pour complémenter les milieux de culture. L'absence de protéines animales dans ce milieu présente un certain nombre d'avantages qui sont discutés