Effect of monovalent rotavirus vaccine on rotavirus disease burden and circulating rotavirus strains among children in Morocco.

Rotarix(TM) vaccine was introduced into the National Program of Immunization of Morocco in October 2010, reaching quickly 87% of the target population of children nationally. The incidence of rotavirus gastroenteritis and the prevalence of circulating rotavirus strains has been monitored in three sentinel hospitals since June 2006. The average percentage of rotavirus positive cases among all children under 5 years old hospitalized for gastroenteritis during the pre-vaccine period (2006-2010) was 44%. This percentage dropped to 29%, 15% and 24% in the 3 years post vaccine introduction (2011, 2012 and 2013), which is a decline of 34%, 66%, and 45%, respectively. Declines in prevalence were greatest among children 0-1 years of age (53%) and were most prominent during the winter and autumn rotavirus season. The prevalence of the G2P[4] and G9P[8] genotype sharply increased in the post vaccine period (2011-2013) compared to the previous seasons (2006-2010). Rotavirus vaccines have reduced greatly the number of children hospitalized due to rotavirus infection at the three sentinel hospitals; it is however unclear if the predominance of G2P[4] and G9P[8] genotypes is related to the vaccine introduction, or if this is attributable to normal genotype fluctuations. Continued surveillance will be pivotal to answer this question in the future.

Diversity of rotavirus strains circulating in children under 5 years of age admitted to hospital for acute gastroenteritis in Morocco, June 2006 to May 2009.

Rotavirus vaccine was introduced in Morocco during 2010. In anticipation of introducing rotavirus vaccines, the Ministry of Health in Morocco established a rotavirus surveillance network in June 2006 at four hospitals in Morocco to obtain baseline data on rotavirus disease burden and prevalent strains. From June 2006 to May 2009, stool samples were collected from children under 5 years of age admitted for diarrhea to four sentinel hospitals serving different regions of Morocco. Rotaviruses were detected in stools using enzyme immunoassay, then genotyped by reverse-transcriptase polymerase chain reaction. Samples with adequate stool in which the P or G types could not be determined by RT-PCR were subjected to nucleotide sequence analysis. Overall, 42% (579 of 1,388) of the stools samples tested were positive for rotavirus. Genotyping of 548 (95%) samples demonstrated that G1P[8] (55%) was the most prevalent strain, followed by G9P[8] (11.3%), G2P[4] (9.1%), G4P[8] (0.9%), and G3P[8] (0.4%). Several other strains were identified including G1P[4] (0.2%), G1P[6] (0.9%), G2P[6] (4.3%), G2P[8] (0.2%), G3P[6] (0.4%), G3P[4] (0.2%), and G9P[6] (0.2%). A high prevalence of mixed infections was found (15% of all samples) of which G1G2P[8] (4%) and G1G3P[8] (3.6%) accounted for the majority. Considerable diversity of rotavirus genotypes was present among strains circulating in Morocco prior to the introduction of the vaccine. This study highlighted the need for maintaining active surveillance to monitor changes in rotavirus disease burden and strain dynamics and to detect changes over time that could impact the effectiveness of the vaccination program.

Comparison of a DNA Based PCR Approach with Conventional Methods for the Detection of Mycobacterium tuberculosis in Morocco.

BACKGROUND: Worldwide, tuberculosis (TB) is a major public health problem and the rapid diagnosis and appropriate chemotherapy become the first priority and a serious challenge to improve TB treatment. In the objective of early TB diagnosis and rapid detection of Mycobacterium tuberculosis (MTB) in the clinical specimens, the utility of the Polymerase Chain Reaction (PCR) using the Insertion Sequence 6110 "IS6110" as target was compared to conventional methods. METHODS: Out of 305 patients with different clinical manifestations: suspected, new, drug relapse, drug failure and chronic cases were enrolled in this study and tested by mycobacteriological and PCR techniques for the investigation about the tubercle bacilli. RESULTS: The results of the in house "IS6110" PCR showed a good sensitivity (92.4%) and high specificity (98.0%), the positive and negative predictive values were 96.4 % and 95.3 % respectively. CONCLUSION: This study showed clearly that the PCR testing using the "IS6110" in the routine analysis is a potential tool for the rapid TB diagnosis, especially for critical cases and would be of great interest to help the clinician in the misdiagnosed critical cases by the traditional radiology.

Evaluation of conventional molecular diagnosis of Mycobacterium tuberculosis in clinical specimens from Morocco.

INTRODUCTION: Tuberculosis is a major public health threat, annually affecting new individuals worldwide, especially those in developing countries. Rapid detection of the agent and effective treatment are two important factors in controlling this disease. METHODOLOGY: The present study aimed to evaluate polymerase chain reaction (PCR) as a rapid and direct molecular method for the diagnosis of Mycobacterium tuberculosis (MTB) in 70 clinical specimens (62 sputum samples, six cerebrospinal fluids, and two biopsies) using heat shock protein (hsp65) as the gene target. Automated sequencing of the same gene was used for the identification of MTB to the species level. RESULTS: The sensitivity of PCR was 81.13%, with specificity of 88.24%; the positive and negative predictive values were 95.56% and 60%, respectively. CONCLUSION: Based on these results, the hsp65 gene sequence can be used to differentiate the members of MTB complex from non-tuberculosis mycobacteria (NTM).

Epidemiology of rotavirus gastroenteritis among children <5 years of age in Morocco during 1 year of sentinel hospital surveillance, June 2006-May 2007.

BACKGROUND: In anticipation of vaccine introduction, we assessed the epidemiology, burden, and genotype of infecting strains of rotavirus disease among Moroccan children hospitalized for acute gastroenteritis. METHODS: From June 2006 through May 2007, 345 children <5 years of age who had acute gastroenteritis and were admitted to 4 sentinel hospitals in different regions of Morocco were enrolled in this surveillance study, and stool specimens were tested for the presence of rotavirus with use of enzyme immunoassay. RNA from positive samples was genotyped by reverse-transcriptase polymerase chain reaction. RESULTS: Overall, 314 children had complete data available, and among these, 138 (44%) tested positive for rotavirus. Rotavirus infection was most common among children <24 months of age (95% of all hospitalizations for rotavirus infection). Rotavirus infection was detected year-round at all 4 sites but was most prevalent from September through January. Genotype analysis demonstrated that 30.6% of samples were G1[P8], 26% were G9[P8], 7.5% were G2[P6], 3.7% were G1[P6], and 0.7% were G2[P8]. Nucleotide sequencing analysis of G- or P-untypeable strains showed that 4.5% were G9[P8], 2.2% were G1[8], 2.2% were G2[P6], and 1.5% were G2[P4]. A high frequency of mixed infection (21%) was found, of which G1G2[P8] accounted for the majority (16.4%). CONCLUSIONS: Rotavirus was responsible for 44% of all hospitalizations for diarrhea among young children at these 4 separate sites in Morocco. These data will help inform a decision on the introduction of rotavirus vaccine in Morocco. Continued and extended surveillance in Morocco will be important to monitor changes in the epidemiology of rotavirus disease and the impact of vaccination after introduction.

Drinking water nitrate and prevalence of methemoglobinemia among infants and children aged 1-7 years in Moroccan areas.

CONTEXT: Nitrate is ubiquitous in environmental media (air, water and soil) and other sources (some medicines, inorganic fertilizers and household's chemicals). It is a hemoglobin-oxidizing agent that can cause methemoglobinemia. The effect of nitrate on infants is well known but less is known about nitrate-induced methemoglobinemia in young children. METHOD: Two cross-sectional studies were carried out in Salé, Morocco to determine the prevalence of methemoglobinemia among 411 infants and children aged 1-7 years in two adjacent areas that were similar in terms of the air quality, available vegetables and medicines but different in terms of the drinking water quality (nitrate-contaminated well water versus municipal water). RESULTS: In the exposed area, nitrate concentration was measured in 78 wells and ranged from 15.39 to 246.90mg/l as NO3-. Nitrate levels were higher than 50mg/l in 69.2% of the surveyed wells, and 64.2% of the participants were drinking nitrate contaminated well waters. The prevalence of methemoglobinemia among study children was 36.2% in the exposed area, and 27.4% in the non-exposed area. Study children drinking well water with a nitrate concentration >50mg/l were significantly more likely to have methemoglobinemia than those drinking well water with a nitrate concentration <50mg/l (p=0.001 at 95% CI=[1.22-2.64]) or than those drinking municipal water (p<0.01 at 95% CI=[1.16-2.21]). In the exposed area, the mean methemoglobin (MetHb) level increased with age (R2= 0.79, p=0.04), whereas in the unexposed area, the mean MetHb level remained relatively stable in the first 6 years of life (R2=0.21, p=0.44). Mean MetHb was normal when the nitrate concentration in water was below 50mg/l as NO3-, and reached an abnormal level, when the nitrate concentration in water ranged between 50 and 90mg/l as NO3-. This last level was statistically similar to mean MetHb at nitrate level above 90mg/l as NO3- (up to 246.9mg/l as NO3-). No association was observed between methemoglobinemia prevalence and gender. This is the first study about methemoglobinemia conducted in Morocco.

Rapid diversification of measles virus genotypes circulating in Morocco during 2004-2005 epidemics.

Measles virus strains circulating in six different regions in Morocco during 2004-2005 were analysed. They were genotyped using two different methods: the recently developed method based on real-time PCR amplification and melting curve analyses, and the conventional method based on nucleic acid sequencing and phylogenetic analysis of 456 nucleotides of the 3'-region of the nucleoprotein (N) gene sequence. Five genotypes (A, B3.2, C2, D7 and D8) were shown to be circulating during this period. Previous studies on measles virus genotypes in Morocco (1998-2003) showed that only the genotype C2 was present and was considered to be endemic. Sequence comparison of the 2004-2005 viruses with other measles strains suggests that measles strains belonging to genotype B3.2 were probably imported from West Africa, whereas those belonging to genotypes D7 and D8 were imported from Europe. These studies which identify the route of importation of measles are important for developing strategies for measles elimination in Morocco.

Genetic analysis of measles viruses isolated in Morocco.

Sequence analysis was conducted on the hemagglutinin (H) and nucleoprotein (N) genes from nine wild-type measles viruses (MV) isolated during an outbreak that occurred in Morocco during 1998 and 1999. The sequence data showed that all the viruses were closely related to each other and were members of genotype C2. Genotype C2 has been shown to be circulating in Europe, and the sequences of the Moroccan isolates were most closely related to the sequences of recent viral isolates from western Europe. This report presents the first molecular epidemiological study of circulating wild-type measles viruses in Morocco. Knowledge of the indigenous strain of measles virus circulating in Morocco will help to describe viral transmission pathways and should contribute to efforts to evaluate the effectiveness of future vaccination campaigns.