Implementation and evaluation of the presto combined qualitative real-time assay for Chlamydia trachomatis and Neisseria gonorrhoeae in Rwanda

Background: The Presto combined qualitative real-time assay for Chlamydia trachomatis and Neisseria gonorrhoeae (Presto CT/NG PCR assay) is appealing for developing countries, because it can be used with multiple DNA extraction methods and polymerase chain reaction (PCR) platforms.Objectives: The objective of the study was to implement and evaluate the Presto CT/NG PCR assay at the National Reference Laboratory (NRL) in Kigali, Rwanda, where no real-time PCR assays for the detection of C. trachomatis or N. gonorrhoeae were available.Methods: The Presto CT/NG PCR assay was first evaluated at the Institute of Tropical Medicine (ITM) in Antwerp, Belgium. Next, NRL laboratory technicians were trained to use the assay on their ABI PRISM 7500 real-time PCR instrument and their competencies were assessed prior to trial initiation. During the trial, endocervical swabs were tested at the NRL, with bi-monthly external quality control testing monitored by the ITM. The final NRL results were evaluated against extended gold standard testing at the ITM, consisting of the Abbott m2000 RealTime System with confirmation of positive results by an in-house real-time PCR assay for C. trachomatis or N. gonorrhoeae.Results: Of the 192 samples analysed using the Presto assay at the NRL, 16 samples tested positive for C. trachomatis and 17 tested positive for N. gonorrhoeae; four of these were infected with both. The sensitivity and specificity of the Presto assay were 93.3% (95% confidence interval [CI]: 68.1% ­ 99.8%) and 99.4% (95% CI: 96.8% ­ 100%) for C. trachomatis and 100% (95% CI: 76.8% ­ 100%) and 98.8% (95% CI: 95.8% ­ 99.9%) for N. gonorrhoeae.Conclusion: C. trachomatis and N. gonorrhoeae testing with the Presto assay was feasible in Kigali, Rwanda, and good performance was achieved

Aspergillus monitoring project in a large educational hospital using molecular assay

Background: It is important to find reliable and accessible methods for the diagnosis and identification of fungal species causing hospital acquired infections. Our main objective was using a rapid and accessible molecular method for the monitoring of Aspergillus infections and identification of causing agents in the level of species. Material and Methods: The study subjects were primarily clinical specimens collected from suspected HAI patients with clinical symptoms after hospitalization. Also some environmental specimens were collected from air and instruments of health care facilities for the investigation of Aspergillus sources in a university hospital of UMSU, Urmia. All specimens were transported to Medical Mycology Center for the detection and identification of Aspergillus species using morphological methods. Also molecular method, PCR-RFLP using single restriction enzyme as a rapid and available method was performed to investigate environmental sources of Aspergillus infections. Results: Total of 110 clinical fungal isolates included Candida and Aspergillus species and some other opportunistic fungi. Among the clinical Aspergillus findings, Aspergillus flavus (47%), Aspergillus fumigatus (29.4%) and Aspergillus niger (23.6%) were the most frequent species respectively and also Aspergillus niger (43.7%), Aspergillus flavus (41.8%), Aspergillus fumigatus (14.7%) were isolated as the most frequent species from environmental sources. Conclusion: Because of accessibility, speed and high sensitivity of diagnosis, the PCR-RFLP was very useful for the identification of medically important Aspergillus species and epidemiological approaches

A Synopsis of Current Malaria Diagnosis Trends

Malaria has remained a major cause of morbidity and mortality in the under developed and developing countries of the tropical and sub-tropical regions of the world. Globally 3.3 billion people live in areas where malaria exists; affecting 300-500 million people annually and it is estimated to be killing approximately 1-3 million people each year and 90of these mortalities occur in African children especially in sub Saharan Africa. Currently; although several control methods are beginning to result in downward trends in incidence in some countries; the gross number of malaria cases is still on the increase due to several factors including poor and ineffective diagnosis. Prompt and effective diagnosis is essential for the management and control of malaria. Over the years evidence has shown that traditional methods for diagnosing malaria remain problematic with a number of limitations. In this synoptic review an update of malaria diagnosis is presented and discussed highlighting the limitations and difficulties of both clinical (symptoms/ clinical signs-based) and laboratory (parasite-based) diagnosis of malaria. Enhancement of accurate malaria diagnosis is now more imperative than ever not only in the background of the current new era of malaria treatment with relatively expensive artemisinin-based combination therapies (ACTs); but more so in the heightened global campaign to effectively control; manage and possibly eradicate malaria from the face of the globe

The impact of the roll-out of rapid molecular diagnostic testing for tuberculosis on empirical treatment in Cape Town, South Africa

Objective To investigate the impact of introducing a rapid test as the first-line diagnostic test for drug-sensitive tuberculosis in Cape Town, South Africa. Methods Xpert® MTB/RIF (Xpert®), an automated polymerase-chain-reaction-based assay, was rolled out between 2011 and 2013. Data were available on 102 007 adults treated for pulmonary tuberculosis between 2010 and 2014. Tuberculosis notification rates per 100 000 population were calculated for each calendar year and for each year relative to the test roll-out locally, overall and by bacteriological confirmation. Empirical treatment was defined as treatment given without bacteriological confirmation by Xpert®, sputum smear microscopy or sputum culture. Findings. Between 2010 and 2014, the proportion of human immunodeficiency virus (HIV)-negative patients treated empirically for tuberculosis declined from 23% (2445/10 643) to 11% (1149/10 089); in HIV-positive patients, it declined from 42% (4229/9985) to 27% (2364/8823). The overall tuberculosis notification rate decreased by 12% and 19% among HIV-negative and HIV-positive patients, respectively; the rate of bacteriologically confirmed cases increased by 1% and 3%, respectively; and the rate of empirical treatment decreased by 56% and 49%, respectively. These changes occurred gradually following the test's introduction and stabilized after 3 years. Conclusion Roll-out of the rapid test in a setting with a high prevalence of pulmonary tuberculosis and HIV infection was associated with a halving of empirical treatment that occurred gradually after the test's introduction, possibly reflecting the time needed for full implementation. More than a quarter of HIV-positive patients with tuberculosis were still treated empirically, highlighting the diagnostic challenge in these patients