Evaluation of the Roche cobas MTB and MTB-RIF/INH Assays in Samples from Germany and Sierra Leone.

The Roche cobas MTB and MTB-RIF/INH assays allow for detection of Mycobacterium tuberculosis complex (MTBC) nucleic acid and rifampicin (RIF) and isoniazid (INH) resistance-associated mutations in an automated, high-throughput workflow. In this study, we evaluated the performance of these assays, employing samples from settings of low and high tuberculosis (TB) burdens. A total of 325 frozen, leftover respiratory samples collected from treatment-naive patients with presumptive TB in Germany (n = 280) and presumptive RIF-resistant TB in Sierra Leone (n = 45) were used in this study. cobas MTB results for detection of MTBC DNA from N-acetyl-l-cysteine-sodium hydroxide (NALC-NaOH)-treated samples were compared to culture results. Predictions of RIF and INH resistance by the cobas MTB-RIF/INH assay were compared to a composite reference standard (phenotypic drug susceptibility testing and line probe assay). Whole-genome sequencing was used to resolve discordances. The overall sensitivity of cobas MTB for detection of MTBC DNA in culture-positive samples (n = 102) was 89.2% (95% confidence interval [CI], 81.7 to 93.9%). The specificity of cobas MTB was 98.6% (95% CI, 96.1 to 99.5%). Sensitivity and specificity for detection of RIF and INH resistance were 88.4% (95% CI, 75.5 to 94.9%) and 97.6% (95% CI, 87.4 to 99.6%) and 76.6% (95% CI, 62.8 to 86.4%) and 100.0% (95% CI, 90.8 to 100.0%), respectively. Discordant results for RIF and INH resistance were mainly due to uncommon mutations in samples from Sierra Leone that were not covered by the cobas MTB-RIF/INH assay. In conclusion, cobas MTB and MTB-RIF/INH assays provide accurate detection of MTBC DNA and resistance-associated mutations in respiratory samples. The influence of regional variations in the prevalence of resistance-conferring mutations requires further investigation

Two Strategies for the Delivery of IPTc in an Area of Seasonal Malaria Transmission in The Gambia: A Randomised Controlled Trial

Bojang and colleagues report a randomized trial showing that delivery of intermittent preventive treatment for malaria in children by village health workers is more effective than delivery by reproductive and child health trekking clinics

South African Ebola diagnostic response in Sierra Leone : a modular high biosafety field laboratory

BACKGROUND : In August 2014, the National Institute for Communicable Diseases (NICD) in South Africa established a modular high-biosafety field Ebola diagnostic laboratory (SA FEDL) near Freetown, Sierra Leone in response to the rapidly increasing number of Ebola virus disease (EVD) cases. METHODS AND FINDINGS : The SA FEDL operated in the Western Area of Sierra Leone, which remained a ªhotspotº of the EVD epidemic for months. The FEDL was the only diagnostic capacity available to respond to the overwhelming demand for rapid EVD laboratory diagnosis for several weeks in the initial stages of the EVD crisis in the capital of Sierra Leone. Furthermore, the NICD set out to establish local capacity amongst Sierra Leonean nationals in all aspects of the FEDL functions from the outset. This led to the successful hand-over of the FEDL to the Sierra Leone Ministry of Health and Sanitation in March 2015. Between 25 August 2014 and 22 June 2016, the laboratory tested 11,250 specimens mostly from the Western Urban and Western Rural regions of Sierra Leone, of which 2,379 (21.14%) tested positive for Ebola virus RNA. CONCLUSIONS : he bio-safety standards and the portability of the SA FEDL, offered a cost-effective and practical alternative for the rapid deployment of a field-operated high biocontainment facility. The SA FEDL teams demonstrated that it is highly beneficial to train the national staff in the course of formidable disease outbreak and accomplished their full integration into all operational and diagnostic aspects of the laboratory. This initiative contributed to the international efforts in bringing the EVD outbreak under control in Sierra Leone, as well as capacitating local African scientists and technologists to respond to diagnostic needs that might be required in future outbreaks of highly contagious pathogens.S1 Video. ªHotº processing of Ebola clinical specimens, PPE and decontamination procedures in South African modular, field-operated biocontainment facility in Sierra Leone.Janusz T Paweska was supported by funding from National Research Foundation and the Global Disease Detection Programmehttp://www.plosntds.orgam2017Microbiology and Plant Patholog

Layout of the SA FEDL in Freetown-Lakka, Sierra Leone with emergency generator and wiring to allow for rapid switch to generator mode in case of power failure.

<p>(<b>A</b>) Biocontainment negative pressure chamber (IsoArk), (<b>B</b>) Room housing biocontainment negative pressure chamber, (<b>C</b>) Donning room, (<b>D</b>) Doffing room, (<b>E</b>) Laboratory airlock area, (<b>F</b>) PCR amplification room, (<b>G</b>) PCR master mix room, (<b>H</b>) Specimens and reagents storage area, (<b>I</b>) RNA extraction room, (<b>J</b>) Facility entrance, (<b>K</b>) Toilet, (<b>L</b>) Office 1, (<b>M</b>) Office 2, (<b>N</b>) Office 3. Petrol generator (5.5 kVa) placement indicated by the red rectangle, distribution of extension cords are indicated with red lines, and emergency connection points by red stars.</p

Number of specimens from EVD suspected cases in Sierra Leone tested daily (total blood and buccal swabs) by SA FEDL during the first weeks of operation, 25 August—30 Sept 2014.

<p>Column = Number of specimens tested daily; Dotted line = Trend line of a number of specimens tested; Solid line = Maximum testing capacity of 58 specimens per day.</p

Ebola virus polymerase gene TaqMan real-time RT-PCR results in blood specimens.

<p>Ebola virus polymerase gene TaqMan real-time RT-PCR results in blood specimens.</p

Operators dressed in BSL3 PPE (scrubs, Tyvek suits, surgical gown, double pair surgical gloves, gumboots, overshoes and PAPR with full face hood) entering the IsoArk main chamber through the airlock.

<p>Operators dressed in BSL3 PPE (scrubs, Tyvek suits, surgical gown, double pair surgical gloves, gumboots, overshoes and PAPR with full face hood) entering the IsoArk main chamber through the airlock.</p

Results of EBOV L-gene RT-PCR in sera subjected to RNA manual (black dots) and automated (circles) extraction.

<p>Results of EBOV L-gene RT-PCR in sera subjected to RNA manual (black dots) and automated (circles) extraction.</p