Prevalensi defisiensi Glucose-6-Phosphate Dehydrogenase (G6PD) pada anak Sekolah Dasar yang tinggal di daerah endemis malaria di Sulawesi utara

Deficiency of glucose-6-phosphate dehydrogenase (G6PD) is the most common enzyme disorder in the world with a very high incidence in the tropics and sub-tropics as a result of malaria selection. North Sulawesi is a part of Indonesian archipelago where malaria has been endemic. This study was aimed to examine the prevalence of G6PD deficiency in the region. An observational cross sectional study was conducted on primary school students belong to different ethnic groups. The purposive sampling method was used to select 442 study subjects, age 5-9 years. The G6PDdeficiency screening test was carried out using G6PD-assay kit. The prevalences of G6PDdeficiency male students were 0% in the Minahasans, 7,4%-12,0% in the Sangihenese, and 4,0%10,3% in the Bolaang Mongondownese. The results suggest that the highest prevalence of G6PD deficiency was in the Sangihe ethnic group. Further molecular analysis would be beneficial to study the genetic relationship of those populations with other neighboring population

Fatal bacteremia due to immotile Vibrio cholerae serogroup O21 in Vientiane, Laos – a case report

<p>Abstract</p> <p>Background</p> <p>Human infections with non-O1, non-O139 <it>V. cholerae </it>have been described from Laos. Elsewhere, non cholera-toxin producing, non-O1, non-O139 <it>V. cholerae </it>have been described from blood cultures and ascitic fluid, although they are exceedingly rare isolates.</p> <p>Case presentation</p> <p>We describe a farmer who died with <it>Vibrio cholerae </it>O21 bacteremia and peritonitis in Vientiane, Laos, after eating partially cooked apple snails (<it>Pomacea canaliculata</it>) and mussels (<it>Ligumia </it>species). The cultured <it>V. cholerae </it>were non-motile. PCR detected <it>ompW </it>and <it>toxR </it>gene regions but not the <it>ctxA, ompU, omp K </it>and <it>TCP </it>gene regions. Although the organisms lacked flagellae on scanning electron microscopy, they possessed the <it>Vibrio </it>flagellin <it>flaA </it>gene.</p> <p>Conclusion</p> <p>Severe bacteremic non-O1, non-O139 <it>V. cholerae </it>is reported from Laos. The organisms were unusual in being non-motile. They possessed the <it>Vibrio </it>flagellin <it>flaA </it>gene. Further research to determine the reasons for the non-motility and virulence is required.</p

A novel PCR-based system for the detection of four species of human malaria parasites and Plasmodium knowlesi.

A microscopy-based diagnosis is the gold standard for the detection and identification of malaria parasites in a patient's blood. However, the detection of cases involving a low number of parasites and the differentiation of species sometimes requires a skilled microscopist. Although PCR-based diagnostic methods are already known to be very powerful tools, the time required to apply such methods is still much longer in comparison to traditional microscopic observation. Thus, improvements to PCR systems are sought to facilitate the more rapid and accurate detection of human malaria parasites Plasmodium falciparum, P. vivax, P. ovale, and P. malariae, as well as P. knowlesi, which is a simian malaria parasite that is currently widely distributed in Southeast Asia. A nested PCR that targets the small subunit ribosomal RNA genes of malaria parasites was performed using a "fast PCR enzyme". In the first PCR, universal primers for all parasite species were used. In the second PCR, inner-specific primers, which targeted sequences from P. falciparum, P. vivax, P. ovale, P. malariae, and P. knowlesi, were used. The PCR reaction time was reduced with the use of the "fast PCR enzyme", with only 65 minutes required to perform the first and second PCRs. The specific primers only reacted with the sequences of their targeted parasite species and never cross-reacted with sequences from other species under the defined PCR conditions. The diagnoses of 36 clinical samples that were obtained using this new PCR system were highly consistent with the microscopic diagnoses

Water, Livelihood and Health in Attapeu Province in Lao PDR

This paper presents the results of an investigation on water-borne infectious diseaseconducted among the people of Attapeu province from 2003 to 2008. Regardless of the lastcholera epidemic in Attapeu province, Lao PDR in the year 1999, the local peoples'awareness of cholera was remarkably low, as demonstrated by the knowledge survey ondiarrheal diseases performed in the province in 2006. In the case study material, derivedfrom continuous field observations on malaria among permanent residents in relocatedvillages in Sanxay district from 2004 to 2008, the infection rate among febrile cases was ashigh as 45% in the early resettlement period, while it was proved that the rate fell later to1.9-14%. Judging from the environmental condition of this settlement area, this papermakes clear the persistent threat of malaria. Furthermore, among the villagers, hookworminfection was highly prevalent. However, liver fluke infections were scarce and noascariasis was found from parasitic stool examination in 2007. Water quality analysis ofthe water sources resulted in remarkably safe water from tube wells from 2003 to 2008

A novel PCR-based system for the detection of four species of human malaria parasites and <i>Plasmodium knowlesi</i>

<div><p>A microscopy-based diagnosis is the gold standard for the detection and identification of malaria parasites in a patient’s blood. However, the detection of cases involving a low number of parasites and the differentiation of species sometimes requires a skilled microscopist. Although PCR-based diagnostic methods are already known to be very powerful tools, the time required to apply such methods is still much longer in comparison to traditional microscopic observation. Thus, improvements to PCR systems are sought to facilitate the more rapid and accurate detection of human malaria parasites <i>Plasmodium falciparum</i>, <i>P</i>. <i>vivax</i>, <i>P</i>. <i>ovale</i>, and <i>P</i>. <i>malariae</i>, as well as <i>P</i>. <i>knowlesi</i>, which is a simian malaria parasite that is currently widely distributed in Southeast Asia. A nested PCR that targets the small subunit ribosomal RNA genes of malaria parasites was performed using a “fast PCR enzyme”. In the first PCR, universal primers for all parasite species were used. In the second PCR, inner-specific primers, which targeted sequences from <i>P</i>. <i>falciparum</i>, <i>P</i>. <i>vivax</i>, <i>P</i>. <i>ovale</i>, <i>P</i>. <i>malariae</i>, and <i>P</i>. <i>knowlesi</i>, were used. The PCR reaction time was reduced with the use of the “fast PCR enzyme”, with only 65 minutes required to perform the first and second PCRs. The specific primers only reacted with the sequences of their targeted parasite species and never cross-reacted with sequences from other species under the defined PCR conditions. The diagnoses of 36 clinical samples that were obtained using this new PCR system were highly consistent with the microscopic diagnoses.</p></div

The second PCR-targeted region of the SSU rRNA genes from variant parasite isotypes.

<p><b>(A)</b> Partial sequences of the variant SSU rRNA genes from malaria parasites. Pf, Pv, Po, and Pm indicate <i>P</i>. <i>falciparum</i>, <i>P</i>. <i>vivax</i>, <i>P</i>. <i>ovale</i> and <i>P</i>. <i>malariae</i> respectively. Pf-standard: The sequence from <i>P</i>. <i>falciparum</i> [GenBank: M19172]. Pf isotype-1 and isotype-2: The identified variant sequences from <i>P</i>. <i>falciparum</i> [Genbank: KJ170099.1 and JQ627151.1]. Pv-standard: The sequence from <i>P</i>. <i>vivax</i> [GenBank: X13926]. Pv isotype-1, isotype-2 and isotype-3: The identified variant sequences from <i>P</i>. <i>vivax</i> [Genbank: U83877.1, KC750244.1 and AF145335.1]. Poc standard: The sequence from <i>P</i>. <i>ovale curtisi</i> [GenBank: L48986]. Poc isotype-1, isotype-2, isotype-3 and isotype-4: The identified variant sequences from <i>P</i>. <i>ovale curtisi</i> [Genbank: KF696376.1, KC633228.1, KJ871671.1 and KF696371.1]. Pow standard: The sequence from <i>P</i>. <i>ovale wallikeri</i> [GenBank: AB182491] Pm standard: The sequence from <i>P</i>. <i>malariae</i> [GenBank: M54897]. Pm isotype-1 and isotype-2: The identified variant sequences from <i>P</i>. <i>malariae</i> [Genbank: KJ619947.1 and KJ170106.1]. All variant sequences from each species are indicated as multiple alignment comparisons. The universal P1 primer region is highlighted in yellow (P1). The inner-specific primers (F2, V3, Oc4 and M4,) are highlighted in blue. The nucleotide changes identified in each variant are highlighted in red. <b>(B)</b> Results of the PCR with the universal primer P1 and the inner-specific primers using the variant DNAs as templates. The PCR reactions were performed with the universal P1 primer and each species-specific primer. The PCR conditions were same as those of the second PCR (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0191886#sec002" target="_blank">Materials and Methods</a>). In each reaction mix, 0.1 ng of the synthesized DNA of each variant sequence was included as template. The products were visualized on 2% agarose-TAE gel containing GelRed (Wako). Lane L indicates a molecular marker (100-bp ladder). The letters shown below each lane indicate the specific primer used for the second PCR reactions (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0191886#pone.0191886.t001" target="_blank">Table 1</a>). Arrows indicate the PCR products (100–106 bp). The template DNAs are indicated below the gels.</p

Determination of the <i>P</i>. <i>falciparum</i> detection limit of the nested PCR system.

<p>The second PCR products were visualized on 2% agarose-TAE gel containing GelRed (Wako). The primer set used for the second PCR was P1 and F2. Lane L indicates a molecular marker (100-bp ladder). The letters shown below each lane indicate the template used for the second PCR reactions (lane P indicates the first PCR products amplified from the sequences of <i>P</i>. <i>falciparum</i>; lanes 10³, 10², 1, 10⁻¹, 10⁻², and 10⁻³ indicate the first PCR products amplified from the DNA extracted from the blood sample containing 10³, 10², 1, 10⁻¹, 10⁻², and 10⁻³ parasites/μL of blood, respectively; lane 0 indicates the first PCR products amplified from the DNA extracted from healthy blood; lane N indicates the diluted first PCR product from water; lane N’, water). The <i>P</i>. <i>falciparum-</i>specific PCR products are indicated with an arrow on the right side.</p

Detailed conditions of the nested PCRs.

<p>Detailed conditions of the nested PCRs.</p