Development of Real-Time PCR Methods for the Detection of Bacterial Meningitis Pathogens without DNA Extraction

Vuong, Jeni; Collard, Jean-Marc; Whaley, Melissa; Issaka, Bassira; Seidou, Issaka; Diarra, Seydou; Ouédraogo, Rasmata; Kambiré, Dinanibè; Taylor, Thomas H.; Sacchi, Cláudio Tavares; Mayer, Leonard W.; Wang, Xin

doi:10.1371/journal.pone.0147765

Cited by 65 publications

(57 citation statements)

References 15 publications

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“…The SDD gene of Haemophilus influenzae, phoE gene of Klebsiella pneumoniae, nucA gene of Staphylococcus aureus, lytA gene of Streptococcus pneumoniae [4], P1 gene of Mycoplasma pneumoniae [5], ctrA gene of Neisseria meningitidis [6] and Toxin gene of Bordetella pertussis [7] were amplified with internal DNA extraction controls T4, as previously described. The SDD gene of Haemophilus influenzae, phoE gene of Klebsiella pneumoniae, nucA gene of Staphylococcus aureus, lytA gene of Streptococcus pneumoniae [4], P1 gene of Mycoplasma pneumoniae [5], ctrA gene of Neisseria meningitidis [6] and Toxin gene of Bordetella pertussis [7] were amplified with internal DNA extraction controls T4, as previously described.…”

Section: Identification Of Respiratory Bacteriamentioning

confidence: 99%

Respiratory and gastrointestinal infections at the 2017 Grand Magal de Touba, Senegal: A prospective cohort survey

Hoang

Goumballa

Dao

et al. 2019

Travel Medicine and Infectious Disease

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Background: The Grand Magal of Touba is the largest Muslim pilgrimage in Senegal with a potential for infectious disease transmission. Methods: Clinical follow-up, adherence to preventive measures and qPCR-based respiratory and gastrointestinal pathogens carriage pre-and post-Magal, were assessed. Results: 110 pilgrims from South Senegal were included. The duration of stay in Touba was 3 days. 41.8% and 14.5% pilgrims reported respiratory and gastrointestinal symptoms. Most individuals having the onset of symptoms during their stay in Touba, or soon after returning. The acquisition of rhinoviruses, coronaviruses and adenovirus was 13.0, 16.7 and 4.6% respectively and that of Streptococcus pneumoniae and Haemophilus influenzae was 3.7% and 26.9%.Acquisition of gastrointestinal viruses and parasites was low, while bacterial acquisition ranged from 2.2% for Campylobacter jejuni to 33.0% for enteropathogenic Escherichia coli. Conclusion: This preliminary study confirms that Grand Magal pilgrims are likely to be exposed to communicable disease risk as observed in other pilgrimage settings. Further study including larger numbers of pilgrims are needed to investigate potential risk factors for respiratory and gastrointestinal infections at the Grand Magal.

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Section: Identification Of Respiratory Bacteriamentioning

confidence: 99%

Respiratory and gastrointestinal infections at the 2017 Grand Magal de Touba, Senegal: A prospective cohort survey

Hoang

Goumballa

Dao

et al. 2019

Travel Medicine and Infectious Disease

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“…No isolates were sent to the Tamale laboratory. PCR assays were used for detection of Streptococcus pneumoniae , Neisseria meningitidis , and Haemophilus influenzae , using gene targets of lytA , sodC , and hpd , respectively [ 11 ]. A cycle threshold (Ct) value ≤36 was considered positive for each bacterium.…”

Section: Methodsmentioning

confidence: 99%

Continued occurrence of serotype 1 pneumococcal meningitis in two regions located in the meningitis belt in Ghana five years after introduction of 13-valent pneumococcal conjugate vaccine

Bozio¹,

Abdul-Karim²,

Abenyeri³

et al. 2018

PLoS ONE

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BackgroundIncreases in pneumococcal meningitis were reported from Ghanaian regions that lie in the meningitis belt in 2016–2017, despite introduction of 13-valent pneumococcal conjugate vaccine (PCV13) in 2012 using a 3-dose schedule (6, 10, and 14 weeks). We describe pneumococcal meningitis epidemiology in the Ghanaian Northern and Upper West regions across two meningitis seasons.MethodsSuspected meningitis cases were identified using World Health Organization standard definitions. Pneumococcal meningitis was confirmed if pneumococcus was the sole pathogen detected by polymerase chain reaction, culture, or latex agglutination in cerebrospinal fluid collected from a person with suspected meningitis during December 2015-March 2017. Pneumococcal serotyping was done using PCR. Annual age-specific pneumococcal meningitis incidence (cases per 100,000 population) was calculated, adjusting for suspected meningitis cases lacking confirmatory testing.FindingsAmong 153 pneumococcal meningitis cases, 137 (89.5%) were serotyped; 100 (73.0%) were PCV13-type, including 85 (62.0%) that were serotype 1, a PCV13-targeted serotype. Persons aged ≥5 years accounted for 96.7% (148/153) of cases. Comparing 2015–2016 and 2016–2017 seasons, the proportion of non-serotype 1 PCV13-type cases decreased from 20.0% (9/45) to 4.1% (3/74) (p = 0.008), whereas the proportion that was serotype 1 was stable (71.1% (32/45) vs. 58.1% (43/74); p = 0.16). Estimated adjusted pneumococcal meningitis incidence was 1.8 in children aged <5 years and ranged from 6.8–10.5 in older children and adults.ConclusionsHigh pneumococcal meningitis incidence with a large proportion of serotype 1 disease in older children and adults suggests infant PCV13 vaccination has not induced herd protection with this schedule in this high-transmission setting.

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“…We used culture or direct real-time PCR testing, or both, for detection of N meningitidis , Haemophilus influenzae , and Streptococcus pneumoniae . 13 Serogroup was deter mined on isolates by slide agglutination and on cerebrospinal fluid specimens that were PCR positive for N meningitidis by serogroup-specific direct real-time PCR. Culture and slide agglutination were also conducted at the Hôpital National de Niamey and a mobile laboratory (LaboMobil, Agence de Médecine Préventive) deployed to a temporary treatment centre in Niamey.…”

Section: Methodsmentioning

confidence: 99%

Emergence of epidemic Neisseria meningitidis serogroup C in Niger, 2015: an analysis of national surveillance data

Sidikou

Zaneidou²,

Alkassoum³

et al. 2016

The Lancet Infectious Diseases

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Summary Background To combat Neisseria meningitidis serogroup A epidemics in the meningitis belt of sub-Saharan Africa, a meningococcal serogroup A conjugate vaccine (MACV) has been progressively rolled out since 2010. We report the first meningitis epidemic in Niger since the nationwide introduction of MACV. Methods We compiled and analysed nationwide case-based meningitis surveillance data in Niger. Cases were confirmed by culture or direct real-time PCR, or both, of cerebrospinal fluid specimens, and whole-genome sequencing was used to characterise isolates. Information on vaccination campaigns was collected by the Niger Ministry of Health and WHO. Findings From Jan 1 to June 30, 2015, 9367 suspected meningitis cases and 549 deaths were reported in Niger. Among 4301 cerebrospinal fluid specimens tested, 1603 (37·3%) were positive for a bacterial pathogen, including 1147 (71·5%) that were positive for N meningitidis serogroup C (NmC). Whole-genome sequencing of 77 NmC isolates revealed the strain to be ST-10217. Although vaccination campaigns were limited in scope because of a global vaccine shortage, 1·4 million people were vaccinated from March to June, 2015. Interpretation This epidemic represents the largest global NmC outbreak so far and shows the continued threat of N meningitidis in sub-Saharan Africa. The risk of further regional expansion of this novel clone highlights the need for continued strengthening of case-based surveillance. The availability of an affordable, multivalent conjugate vaccine may be important in future epidemic response.

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Development of Real-Time PCR Methods for the Detection of Bacterial Meningitis Pathogens without DNA Extraction

Cited by 65 publications

References 15 publications

Respiratory and gastrointestinal infections at the 2017 Grand Magal de Touba, Senegal: A prospective cohort survey

Respiratory and gastrointestinal infections at the 2017 Grand Magal de Touba, Senegal: A prospective cohort survey

Continued occurrence of serotype 1 pneumococcal meningitis in two regions located in the meningitis belt in Ghana five years after introduction of 13-valent pneumococcal conjugate vaccine

Emergence of epidemic Neisseria meningitidis serogroup C in Niger, 2015: an analysis of national surveillance data

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