Real-time PCR has been developed to genotype measles virus (MV) isolates. MV strains circulating in epidemics inWe developed a real-time amplification refractory mutation system PCR (RT-AMRS PCR) using SYBR green fluorescent dye. Six pairs of primers for RT-ARMS PCR were designed to specifically amplify genotypes A, B2, B3.1, B3.2, C2, and D7. Genotypes could be differentiated by melting curve analysis. All strains were also confirmed by direct sequencing. Using the result obtained by direct sequencing and phylogenetic analysis as the reference, the accuracy of MV by RT-ARMS PCR and melting curve analysis was 97%. However, the latter method is more rapid and sensitive than the former method. This method could be a useful tool for molecular epidemiological studies of MV, providing an efficient alternative for large-scale studies.Measles virus (MV) infection is a public health problem worldwide, with an estimated 30 million people infected and 530,000 deaths in 2003 (21). MV is serologically a monotypic virus; however, diversity within the complete hemagglutinin (H) gene and the C-terminal part of the nucleoprotein (N) gene has allowed the classification of MV into eight clades, designated A to H, including 22 genotypes (22). Measles has been designated as a target for eradication by the WHO by the year 2010. According to a strategic plan for the elimination of measles, molecular epidemiology will play an essential role in measuring the impact of the vaccination campaign in countries where the disease remains endemic and in tracing the source of MVs isolated in countries with an advanced eradication program, where most cases are imported.The WHO recommends the genotyping of representative strains in every outbreak. This is essentially performed by nucleotidic sequencing of genomic fragments followed by phylogenetic analysis. Although this method is the "gold standard" and allows direct classification, it is time-consuming and expensive and does not allow rapid analysis of numerous samples. Alternative methods have been described, such as restriction fragment length polymorphism and nucleotide-specific multiplex PCR to differentiate MV strains (11,18). These methods rely on gel electrophoreses analysis that can lead to ambiguous interpretations. Samuel et al. (17) developed a method based on a modification of the amplification refractory mutation system, which was limited to a single genotype and needed a post-PCR enzyme-linked immunosorbent assay analysis, which is time-consuming.In this study, a method for genotyping measles virus was performed by real-time amplification refractory mutation system (RT-ARMS) PCR using SYBR green fluorescent dye. The method uses genotype-specific primers to specifically amplify MV genotypes. This novel assay allows real-time detection of PCR products, and the genotype is subsequently determined by melting curve analysis. In the present studies, RT-ARMS PCR was used to genotype 30 isolates from MV outbreaks in Africa and France. The results using this method were confirmed by compar...

Since the confirmation of measles cases represents an important indicator regarding the performance of the measles-elimination program, the aim of this study was to evaluate the effectiveness of the routine procedures followed in Morocco for the laboratory confirmation of measles cases. Suspected cases reported between January 2010 and December 2012 were assessed for the timeliness of the sample collection, occurrence of measles clinical symptoms, and the results of the laboratory diagnoses. For 88% of the 2,708 suspected cases, a clinical specimen was collected within 7d of rash onset, of which 50% were IgM-positive and 2.6% were equivocal. The measles symptoms were reported in 91.4% of the cases; the occurrence of symptoms showed a positive association with the serological results (odds ratio [OR] = 2.9883, 95% confidence interval [CI] 2.2238–4.0157). Of the negative samples, 52% (n = 116) tested positive by real-time polymerase chain reaction (PCR). These results are in favor of using molecular detection to complement serological diagnosis in the context of measles surveillance approach in Morocco. In addition, the introduction of additional laboratory methods for differential diagnosis is required for the final classification of suspected cases with maculopapular rash and fever in the context of the measles elimination program.

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.

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.

Background: Intestinal parasite spread in tropical countries is especially common among primary school students. This study aimed to determine the frequencies of the intestinal parasite by different techniques among school students in Alkalakla locality, Khartoum state. Methods: This study was conducted in school students in Al-kalakla locality in Khartoum state from period between 20th December 2016 to 5th May 2017. Stool samples were collected from 134 randomly selected students, of whom 67 were males and 67 were females.  All samples were examined using the wet preparation technique, formal ether concentration technique and saturated sugar floatation technique. Results: The frequency of intestinal parasites was 35.5% overall in the students examined; females were more affected than males (38.8% and 32.8%, respectively). The more affected age groups were 12-14 years followed by 9-11 and 6-8 years old (53.8%, 36.3% and 26.4% respectively). The least frequent intestinal parasite was Taenia spp. (1.5%) followed by Giardia lamblia (3.7%), Schistosoma mansoni and Ascaris lumbricoides (5.2% each), Entamoeba coli (7.5%), Hymenolepis nana (10.4%), and Entamoeba histolytica (16.4%). In total, 20.9% were infected with single parasite while 14.9% were infected with more than one parasite. The frequency of parasite by formal ether concentration method was 35.8 %, by wet preparation method was 17.9 % and by the saturated sugar flotation method was 16.4%. Conclusion: Our data showed that intestinal parasites were common in school students; however, females were more affected than males and the 12-14-years age group was the most affected age group.  The formal ether concentration method was the best method for detecting of intestinal parasite.