Imported malaria to Makkah district, Saudi Arabia: is there any risk of local transmission?

In Makkah, Saudi Arabia, there is an impending risk of imported malaria. This risk comes from the fact that millions of people, in majority from tropical and subtropical countries where malaria is endemic, visit the country to perform Hajj and Umrah every year. Moreover, millions of expatriates from endemic countries come to Makkah for work. Likewise, many Saudi citizens travel to endemic areas overseas for business and pleasure. We performed a retrospective analysis of all reported malaria cases in Makkah region, Saudi Arabia for years 2014 and 2015. In addition, sorting of mosquito populations in Makkah region was undertaken. Based on national data regarding reported malaria cases, 235 malaria cases were recorded in years 2014 and 2015. Of the reported cases 232 were non-Saudi and only 3 cases were Saudi. Those recorded Saudi cases were just returning from a travel to an endemic area. Most of the cases [79.6%] were P. falciprum and the remaining was P. vivax. Infected male represent 62% and female represent 38%. Age of the majority of reported cases [71.5%] lie between 31 and 50 years. Most of reported cases were from Chad, Pakistan, Nigeria and Sudan. Sorting of mosquito populations revealed the absence of malaria vectors in Makkah District

Assessment of three blood genomic-DNA preparation methods for malaria molecular diagnosis

Species-specific PCR techniques are highly sensitive and reliable alternatives to clSpecies-specific PCR techniques are highly sensitive and reliable alternatives to classical methods for malaria diagnosis and speciation, especially in endemic regions under advanced control or elimination programs where asymptomatic and low-density infections are increasingly reported. Nevertheless, the performance of these techniques is directly affected by the quality of isolated DNA templates. A Plasmodium falciparum/vivax-specific diagnostic Nested- PCR [Pf/Pv N-PCR] was used to assess three DNA preparation methods, Qiagen[registered sign] Mini- Chromatographic kit [QIAmp[registered sign]] and Jena-Biosciences[registered sign]DNA isolation kit [JB[registered sign]] for genomic DNA extraction from EDTA-preserved whole blood samples, and Whatman-FTA[registered sign] purification reagent [FTA[registered sign]] for DNA preparation from dry blood spots [DBS] collected onto FTA[registered sign]- cards

A total of 84 out of 137 blood specimens collected from malaria suspicious febrile patients who visited five health care centres in south-western endemic localities of Saudi Arabia were found P. falciparum positive by at least one method. Among these, only 76 [90%] were reported P. falciparum malaria positive by two expert microscopists. No other species of Plasmodium were detected. Pf/Pv N-PCR revealed 84/84 [100%], 75/84 [89%], and 81 [96%] P. falciparum positive samples using DNA templates prepared by QIAmp[registered sign], JB[registered sign], and FTA[registered sign] purification methods, respectively. Therefore, Pf/Pv N-PCR, when applied to QIAmp[registered sign] DNA templates showed to be a highly sensitive diagnostic method, particularly useful for submicroscopic specimens from clinically malaria suspicious patients in endemic areas. On the other hand, Pf/Pv N-PCR of FTA[registered sign]-DBS DNA templates revealed 5 positive cases missed by microscopy, encouraging its use as an affordable field semi-adapted protocol for malaria active screening, especially in remote rural regions with limited laboratory infrastructure

Appraisal of prenatal anti-Toxoplasma gondii [IGG+IGM]-IHA/IGM-Elisa screening in single samples VIA IGG Avidity test

Congenital toxoplasmosis is associated with important morbidity and mortality. Since vertical transmission of Toxoplasma gondii can occur in acute cases, antenatal screening for recent infections is vital. Accurate determination of acute toxoplasmosis requires a combination of immunoassays, usually not routinely applied for screening purposes. This study evaluated the anti-T. gondii [IgG+IgM]/IgM prenatal screening procedure by IgG avidity assay

The routine prenatal screening for [IgG+IgM] anti-T. gondii by indirect hemagglutination [IHA] in serum samples was done of 2247 pregnant women who attended two hospitals between 2011 and 2013 revealed 487 [21.7%] positive samples. Examination of IHA-positive sera by IgM and IgG/IgG-avidity concurrent ELISA tests revealed 7 positive and 3 border-line IgM-ELISA titers during the initial check-up of 10 women, who were then followed up at 3-4 week-intervals. Among these, 4 [40%] showed simultaneous high avidity IgG antibodies, indicating distant infection by the parasite, and no anti-T. gondii specific IgG could be detected in follow-up sera of two cases [20%], indicating false IgM initial positive results. Only 4 [40%] women showed simultaneous IgM and low avidity IgG antibodies indicating active infections. Avoidance of an overdiagnosis of acute toxoplasmosis Anti-T. gondii [IgG+IgM]/IgM prenatal screening must be supplemented by a discriminative test like IgG avidity ELISA

Comparison of a genus-specific conventional PCR and a species-specific nested-PCR for Malaria diagnosis using FTA collected samples from Kingdom of Saudi Arabia

Molecular tools are increasingly accepted as the most sensitive and reliable techniques for malaria diagnosis and epidemiological surveys. Also, collection of finger prick blood spots onto filter papers is the most simple and affordable method for samples preservation and posterior molecular analysis, especially in rural endemic regions where malaria remains a major health problem. Two malaria molecular diagnostic tests, a Plasmodium genus -specific conventional PCR and a Plasmodium species -specific Nested PCR, were evaluated using DNA templates prepared from Whatman - FTA -cards' dry blood spots using both, Methanol -fixation/Heat -extraction and FTA commercial purification kit. A total of 121 blood sam -ples were collected from six Saudi south -western endemic districts both, as thick and thin films for routine microscopic screening and onto FTA - cards for molecular studies. Out of the 121 samples, 75 were P. falciparum positive by at least one technique. No other species of Plasmodium were detected. P. falciparum parasites were identified in 69/75 [92%] samples by microscopic screening in health care centers. P. genus -specific PCR was able to amplify P. falciparum DNA in 41/75 [55%] and 59/75 [79%] samples using Methanol -fixation/Heat -extraction and FTA purification kit, respectively. P. species -specific Nested PCR revealed 68/75 [91%] and 75/75 [100%] positive samples using DNA templates were isolated by Methanol -fixation/Heat - extraction and FTA purification methods, respectively. The species -specific Nested PCR applied to Whatman -FTA preserved and processed blood samples represents the best alternative to classical microscopy for malaria diagnosis, particularly in epidemiological screening

Malaria drug resistant: current situation with reference to Saudi Arabia [review]

Malaria is a chronic disease caused by parasitic protozoa of plasmodia species. Four plasmodium species are causing malaria to human [P. vivax, P. ovale, P. ma-lariae and P.falciparum]. Malaria classifies as one of the most serious diseases in tropical and subtropical countries and p. falciparum represents the major cause of death by malaria species. Approximately 40% of the world population resides in areas of active malaria transmission. Treatment and prophylaxis measures are important to reduce morbidity and mortality rate of infection. In last two decades, a significant number of malaria drug resistance cases [mainly P. falciparum] were reported in endemic areas against choroquine components. Parasite showed enormous amount of antigenic variation under immune pressure leading to emergence of vaccine resistant strains. Similarly, under drug pressure it allows mutations to settle in the target genes. It is becoming more and clearer that with the continuous exposure to a drug, the parasite accumulates more and more number of mutations in these genes. Artemisinin is the only available drug that is globally effective. This review concentrates on the current situation of malaria drug resistance including epidemiological distribution, the mechanism of how the malaria resists certain drugs and the role of recent advances facilities in molecular biology to evaluate the impact on drug resistance of new drug-based strategies in Saudi Arabia

PCR assay in malaria diagnosis using filter paper samples from Jazan region, Saudi Arabia

PCR assay using designed primers was evaluated in detecting human malaria infection from whole blood and/or on Whatman filter-paper compared to conventional microscopy. Two DNA extraction methods were used for dried blood-spots; QIAamp mini kit and methanol-fixation/heat-extraction. A total of 118 cases were collected from 4 hospitals at Jazan district. Microscopic examination showed positivity in 66/118 samples [56%]. Thin films showed parasitae-mia from 1+ to 4+. In PCR assay, 79 samples [70%] were positive for the genus Plasmodium given 153 base pair PCR product. All microscopy positive samples were PCR positive but PCR detected 13 cases missed by microscopy. As for filter-paper spot samples, 68 samples [57.6%] were PCR positive when DNA was extracted by QIAamp mini kit whereas 49 [41.5%] by methanol-fixation/heat-extracion method. PCR sensitivity decreased by using DNA extracted from filter-paper compared to microscopy and whole blood PCR. But the DNA isolated from filter paper detected parasites in many microscopy negative samples

Preliminary study of the prevalence of intestinal parasites among diarrheic inhabitants in Makkah Al Mukarramah

Diarrheic disease is one of the greatest causes of morbidity and mortality worldwide. Intestinal parasites contribute to the disease and the well being of humans. This study was undertaken in the Holly City of Makkah Al-Mukarramah. A total of 166 diarrheic stool samples were collected. A wet smear from each specimen in normal saline and Lugol's iodine solution was examined microscopically for the trophozoites and cysts of protozoan parasites. Stools were also examined using ethyl acetate formalin concentration technique to confirm the diagnosed parasites. All stool specimens were stained by Ziehl-Neelsen stain to detect the oocysts of Crypto-sporidium spp. One way ANOVA was used to analyze data. 128 persons were found to be infected, with an overall prevalence of 77.1%. 46.99% of the samples were females, and 53.01% were males. The prevalence of infection in females was 36.1%, and 40.9% in males. 16.9% of infected females were living near the Holy Masjid [down town], while 19.3% were living away from the Holy Masjid [up town]. 18.7% of infected males were living down town, while 22.3% were living up town. The majority of cases fall into the young age groups [<30 years old]. There is no significant difference between the prevalence of infection down town and up town [P=0.22], whereas the prevalence of infection between the patients over or under 30 years old was significance [P=0.036]. The rates of infection were higher in those living up town than those living down town. The results were critically discussed

Detection of drug resistance markers for chloroquine and pyrimethamine-sulfadoxine in Jazan area, Saudi Arabia using PCR and restriction digestion

Nested PCR and restriction analysis were used to detect mutations at codon 76 of Plasmodium falciparum chloroquine resistance transporter gene [pfcrf] and codon 59 of dihydrofolate reductase gene [dhfr] that indicate chloroquine [CQ] and pyri-methamine-sulfadoxine [PYR-SDX] resistance respectively. P. falciparum isolates from malaria-endemic area of Jazan showed CQ resistance rate [89.5%], the highest percentage of chloroquine resistance ever recorded in Saudi Arabia. One the other hand, 10.5% of isolates showed a PYR-SDX resistant allele as a first reported in the kingdom. The use of molecular markers as additional tools to map areas of chloroquine resistance was expected to contribute in the development of new strategies for therapeutic intervention towards malaria in Saudi Arabia

Detection malaria in saudi arabia by real time PCR

Malaria transmission occurs in Saudi Arabia and mainly endemic in the lowlands of Asir region, the southwester province. Imported cases have been reported. Sensitive routine laboratory techniques for rapid and accurate malaria diagnosis are therefore desirable to facilitate the identification of individuals infected with the malarial parasites and to follow up the progress of treatment of such cases with appropriate drugs. Traditional diagno sis, based on the microscopic examination of Giemsastained thick and thin films remains the main standard method of diagnosis used for malaria diagnosis in Saudi Arabia. Molecular diagnostic techniques based on the detection of nucleic acid [as PCR; Realtime PCR] are now highly considered. Real time PCR a new methodology has been recently applied to detect human malaria. In this study a total of forty four samples, using wholeblood, dried blood and thick smears were examined by PCR and Realtime PCR. Both techniques showed a higher sensitivity than the microscopy. Parasites were detected in twenty nine samples out of forty four, compared to twenty six of thirty nine were positive with thin blood film. Realtime PCR assay offers a practical and positive alternative for rapid and accurate diagnosis for malaria infection. The application of such technique will be significantly valuable especially for screening for malaria infection in endemic areas.