Comparative evaluation of automated KingFisher Flex Purification System 96 (ThermoFisher Scientific) and manual QIAamp Viral RNA Mini Kit (Qiagen) extraction methods for SARS-CoV-2

Background: The extraction step of the viral material of the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) influences the quality of reverse transcriptase-polymerase chain reaction (RT-PCR) results in diagnosis of coronavirus disease 2019 (COVID-19). The purpose of this cross-sectional study was to evaluate the diagnostic performance of the automated extraction system "KingFisher Flex Purification System 96 (ThermoFisher)" compared to the manual method with the "QIAamp Viral RNA Mini Kit (Qiagen)". Methodology: From October to December 2020, comparative diagnostic evaluation of two methods of SARSCoV-2 RNA extraction methods was conducted on 159 fresh and 120 frozen nasopharyngeal and oropharyngeal specimens collected from travellers and suspected cases or contacts of COVID-19 patients in Burkina Faso. The FastPlexTM Triplex 1-Step COVID 19 Detection Kit (RT-PCR, RNA extraction free) (Precigenome LLC) was used to amplify on the same PCR plate, RNA extracts from manual QIAamp Viral RNA Mini Kit and automated KingFisher Flex Purification System 96 (ThermoFisher) using the QuantStudio5 thermal cycler (Applied Biosystems). Analysis of the diagnostic performance of the SARS-CoV-2 RT-PCR assay following RNA extraction by the two methods was done using an online OpenEpi software. Results: For fresh samples, the study found a slightly higher RT-PCR positivity rate following manual extraction (12.6%) than automated extraction (9.4%). For frozen samples, the positivity rate was far higher for manual (38.33%) than automated extraction method (20.83%). The results show that the performance of the automated extraction was inferior when compared to the manual extraction for both fresh samples (sensitivity 35%, specificity 94.2%) and frozen samples (sensitivity 43.5%, specificity 93.2%). However, using McNemar Chi-square with Yates correction, there was no significant difference in positivity rate of RT-PCR (x2=0.76, p=0.38) between the two extraction methods for the fresh samples, but there was a significant difference (x2=12.9, p= 0.0003) in the extraction of the frozen samples. Conclusion: The results of this study showed that KingFisher Flex Purification System 96 (ThermoFisher) automatic extraction method was less sensitive and specific than QIAamp Viral RNA Mini Kit (Qiagen) manual extraction method. This information can serve as guide to laboratories in the choice of RNA extraction methods to use for RT-PCR detection of SARS-CoV-2

Magnitude and associated factors of latent tuberculosis infection due to Mycobacterium tuberculosis complex among high-risk groups in urban Bobo-Dioulasso, Burkina Faso

Objectives: To determine the prevalence and risk factors for latent tuberculosis infection (LTBI) among three high-risk groups - household contacts of TB index cases, healthcare workers and slaughterhouse workers - in Bobo-Dioulasso, Burkina Faso. Methods: Participants were recruited to this cross-sectional study from March to July 2020 after giving informed consent. Sociodemographic, clinical and biological data were collected using a structured questionnaire. The QuantiFERON-TB Gold Plus test (QFT-Plus) and the tuberculin skin test (TST) were used for detection of LTBI. Bivariate and multivariate logistic regression analyses were performed to identify risk factors for LTBI. Results: The prevalence of LTBI among 101 participants (age range 15-68 years) was 67.33% [95% confidence interval (CI) 57.27-76.33] and 84.16% (95% CI 75.55-90.66) based on QFT-Plus and TST results, respectively. Compared with healthcare workers and household contacts of TB index cases, the prevalence of LTBI among slaughterhouse workers was significantly higher for both QTF-Plus (96.8%; P /=15 years of exposure (AOR 5.617, 95% CI 1.202-32.198), having an animal at home (AOR 2.735, 95% CI 1.102-6.789) and protozoal infection (AOR 2.591, 95% CI 1.034-6.491) were significantly associated with LTBI on the QFT-Plus assay. Conclusion: The prevalence of LTBI was high in all three groups, particularly slaughterhouse workers. The risk factors identified could form the basis of targeted intervention

Molecular diagnosis of COVID-19 in Burkina Faso: successful challenge

COVID-19 has worsened the health situation in Burkina Faso. In fact, the country has known a peak of the second wave, which began in November, and ended around January 2021. Biological diagnosis has played a key role in the management of COVID-19. The aim of this review paper is to address the practical aspects that laboratories have faced in order to meet the challenge of SARS-CoV-2 diagnosis in Burkina Faso. According to international requirements, Burkina Faso has used real-time Reverse Transcription Polymerase Chain Reaction (rRT-PCR) as the “gold standard” for the diagnosis of COVID-19. From March 9, 2020 to July 31, 2021, in Burkina Faso, laboratories involved in COVID-19 diagnosis analyzed 226,189 samples by molecular tests and 2, 352 samples by rapid antigenic tests, whose peak was in January 2021 with 35,984 samples analyzed. The daily average rate of samples analysis was 456.02 tests. The majority of the individuals requesting COVID-19 tests were travelers (62.00%), followed by contact cases (18.42%), suspected cases (7.95%), voluntary screening (7.57%), and 4.06% of other applicants consisting of health care personnel and at-risk patients. In terms of prevention, vaccines are being administered to the general population. However, some efforts must be made to provide automated sample analysis equipment and complete sequencing of SARS-CoV-2 remains among the challenges

Effect of blood type on anti-a-Gal immunity and the incidence of infectious diseases

The identification of factors affecting the susceptibility to infectious diseases is essential toward reducing their burden on the human population. The ABO blood type correlates with susceptibility to malaria and other infectious diseases. Due to the structural similarity between blood antigen B and Gala1-3GalB1-(3)4GlcNAc-R (a-Gal), we hypothesized that self-tolerance to antigen B affects the immune response to a-Gal, which in turn affects the susceptibility to infectious diseases caused by pathogens carrying a-Gal on their surface. Here we found that the incidence of malaria and tuberculosis, caused by pathogens with a-Gal on their surface, positively correlates with the frequency of blood type B in endemic regions. However, the incidence of dengue fever, caused by a pathogen without a-Gal, was not related to the frequency of blood type B in these populations. Furthermore, the incidence of malaria and tuberculosis was negatively correlated with the anti-a-Gal antibody protective response. These results have implications for disease control and prevention.Peer reviewedVeterinary Pathobiolog

Global respiratory syncytial virus–related infant community deaths

Background Respiratory syncytial virus (RSV) is a leading cause of pediatric death, with >99% of mortality occurring in low- and lower middle-income countries. At least half of RSV-related deaths are estimated to occur in the community, but clinical characteristics of this group of children remain poorly characterized. Methods The RSV Global Online Mortality Database (RSV GOLD), a global registry of under-5 children who have died with RSV-related illness, describes clinical characteristics of children dying of RSV through global data sharing. RSV GOLD acts as a collaborative platform for global deaths, including community mortality studies described in this supplement. We aimed to compare the age distribution of infant deaths <6 months occurring in the community with in-hospital. Results We studied 829 RSV-related deaths <1 year of age from 38 developing countries, including 166 community deaths from 12 countries. There were 629 deaths that occurred <6 months, of which 156 (25%) occurred in the community. Among infants who died before 6 months of age, median age at death in the community (1.5 months; IQR: 0.8−3.3) was lower than in-hospital (2.4 months; IQR: 1.5−4.0; P < .0001). The proportion of neonatal deaths was higher in the community (29%, 46/156) than in-hospital (12%, 57/473, P < 0.0001). Conclusions We observed that children in the community die at a younger age. We expect that maternal vaccination or immunoprophylaxis against RSV will have a larger impact on RSV-related mortality in the community than in-hospital. This case series of RSV-related community deaths, made possible through global data sharing, allowed us to assess the potential impact of future RSV vaccines