Evaluation of new multiplex PCR primers for the identification ofPlasmodium species found in Sabah, Malaysia

Stanis, Cheronie Shely; Song, Beng Kah; Chua, Tock Hing; Lau, Yee Ling; Jelip, Jenarun

doi:10.3906/sag-1411-114

Cited by 15 publications

(25 citation statements)

References 40 publications

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“…These results coincided with those from previous work reporting 75–98% sensitivity for PCR regarding the identification of Plasmodium spp. [38, 39, 57, 58], together with 98–100% estimations for detecting P. vivax [38, 59, 60]. Similarly, PCR estimated higher prevalence values for the species evaluated and for certain types of co-infection, such increases having been observed in previous studies for both simple and mixed infections [22, 23, 27, 47, 48].…”

Section: Discussionsupporting

confidence: 60%

Plasmodium malariae in the Colombian Amazon region: you don’t diagnose what you don’t suspect

Niño

Cubides

Camargo-Ayala

et al. 2016

Malar J

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BackgroundMalaria is a worldwide public health problem; parasites from the genus Plasmodium spp. are the aetiological agent of this disease. The parasite is mainly diagnosed by microscope-based techniques. However, these have limited sensitivity. Many asymptomatic infections are sub-microscopic and can only be detected by molecular methods. This study was aimed at comparing nested PCR results to those obtained by microscope for diagnosing malaria and to present epidemiological data regarding malaria in Colombia’s Amazon department.MethodsA total of 1392 blood samples (taken by venepuncture) from symptomatic patients in Colombia’s Amazon department were analysed in parallel by thick blood smear (TBS) test and nested PCR for determining Plasmodium spp. infection and identifying infecting species, such as Plasmodium vivax, Plasmodium malariae and/or Plasmodium falciparum. Descriptive statistics were used for comparing the results from both tests regarding detection of the disease, typing infecting species and their prevalence in the study region. Bearing the microscope assay in mind as gold standard, PCR diagnosis performance was evaluated by statistical indicators.ConclusionThe present study revealed great differences between both diagnostic tests, as well as suggesting high P. malariae prevalence from a molecular perspective. This differed profoundly from previous studies in this region of Colombia, usually based on the TBS test, suggesting that diagnosis by conventional techniques could lead to underestimating the prevalence of certain Plasmodium spp. having high circulation in this area. The present results highlight the need for modifying state malaria surveillance schemes for more efficient strategies regarding the detection of this disease in endemic areas. The importance of PCR as a back-up test in cases of low parasitaemia or mixed infection is also highlighted.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-016-1629-3) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 60%

Plasmodium malariae in the Colombian Amazon region: you don’t diagnose what you don’t suspect

Niño

Cubides

Camargo-Ayala

et al. 2016

Malar J

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“…Multiple sensitive molecular methods for P. knowlesi detection have been published to date including nested [4,22], single-step [27][28][29], and real-time PCR [30,31], and loop-mediated isothermal ampli cation (LAMP) [32][33][34][35]. Although these molecular methods are directed against a range of different P. knowlesi gene targets, their reported detection limits are all below that of routine microscopic examination of malaria blood lms.…”

Section: Discussionmentioning

confidence: 99%

Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia

Nuin¹,

Tan

Lew³

et al. 2020

Preprint

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Background : The monkey parasite Plasmodium knowlesi is an emerging public health issue in Southeast Asia. In Sabah, Malaysia, P. knowlesi is now the dominant cause of human malaria. Molecular detection methods for P. knowlesi are essential for accurate diagnosis and in monitoring progress towards malaria elimination of other Plasmodium species. However, recent commercially available PCR malaria kits have unpublished P. knowlesi gene targets or have not been evaluated against clinical samples. Methods : Two real-time PCR methods currently used in Sabah for confirmatory malaria diagnosis and surveillance reporting were evaluated: the QuantiFast™ Multiplex PCR kit (Qiagen, Germany) targeting the P. knowlesi 18S SSU rRNA; and the abTES™ Malaria 5 qPCR II kit (AITbiotech, Singapore), with an undisclosed P. knowlesi gene target. Diagnostic accuracy was evaluated using 52 P. knowlesi, 25 P. vivax , 21 P. falciparum , and 10 P. malariae clinical isolates, and 26 malaria negative controls, and compared against a validated reference nested PCR assay. The limit of detection (LOD) for each PCR method and Plasmodium species was also evaluated. Results : The sensitivity of the QuantiFast™ and abTES™ assays for detecting P. knowlesi was comparable at 98.1% (95%CI 89.7-100) and 100% (95%CI 93.2-100) respectively. Specificity of the QuantiFast™ and abTES™ for P. knowlesi was high at 98.8% (95%CI 93.4-100) for both assays. The QuantiFast™ assay demonstrated falsely-positive mixed Plasmodium species at low parasitaemias in both the primary and LOD analysis. Diagnostic accuracy of both PCR kits for detecting P. vivax , P. falciparum , and P. malariae was comparable to P. knowlesi . The abTES™ assay demonstrated a lower LOD for P. knowlesi of ≤0.125 parasites/µL compared to QuantiFast™ with a LOD of 20 parasites/µL. Hospital microscopy demonstrated a sensitivity of 78.8% (95%CI 65.3-88.9) and specificity of 80.4% (95%CI 67.6-89.8) compared to reference PCR for detecting P. knowlesi . Conclusion : The QuantiFast™ and abTES™ commercial PCR kits performed well for the accurate detection of P. knowlesi infections. Although the QuantiFast™ kit is cheaper, the abTES™ kit demonstrated a lower LOD, supporting its use as a second-line referral-laboratory diagnostic tool in Sabah, Malaysia.

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“…namely P. malariae or P. knowlesi. Furthermore, of the 28 studies, ten studies (12,24,25,28,31,34,41,52,61,63) conducted PCR in the whole samples regardless of the microscopy results in order to trace the sub-microscopic infections. In addition, 18 studies utilised only one method of detection for malaria.…”

Section: Description Of Included Studiesmentioning

confidence: 99%

“…However, RDTs do not provide parasite quanti cation and are considered more expensive than light microscopy (19). Nevertheless, the advent of molecular techniques such as PCR are more accurate in identi cation and differentiation of all malaria species than microscopy and RDT (10,(27)(28)(29). Despite the greater sensitivity of PCR, it is not convenient for eld and resource-limited settings due to the requirement of complex equipment, reagents and know-how (19).…”

Section: Introductionmentioning

confidence: 99%

Malaria distribution and performance of malaria diagnostic methods in Malaysia (1980-2019): a systematic review

Rahim

Munajat

Idris

2020

Preprint

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Background: Malaysia has already achieved remarkable accomplishments in reaching zero indigenous human malaria cases in 2018. Prompt malaria diagnosis, surveillance and treatment played a key role in the country’s elimination success. Looking at the dynamics of malaria distribution during the last decades might provide important information regarding the potential challenges of such an elimination strategy. This study was performed to gather all data available in term of prevalence or incidence on Plasmodium infections in Malaysia over the last four decades. Methods: A systematic review of the published English literature was conducted to identify malaria distribution from 1980 to June 2019 in Malaysia. Two investigators independently extracted data from PubMed, Scopus, Web of Science and Elsevier databases for original papers.Results: The review identified 46 epidemiological studies in Malaysia over the 39-year study period, on which sufficient information was available. The majority of studies were conducted in Malaysia Borneo (31/46; 67.4%), followed by Peninsular Malaysia (13/46; 28.3%) and in both areas (2/46; 4.3%). More than half of all studies (28/46; 60.9%) were assessed by both microscopy and PCR. Furthermore, there was a clear trend of decreases of all human malaria species with increasing P. knowlesi cases throughout the year of sampling period. The summary estimates of sensitivity were higher for P. knowlesi than other malaria species for both microscopy and PCR. Nevertheless, the specificities of summary estimates were similar for microscopy (40 – 43%) but varied for PCR (2 – 34%).Conclusions: This study outlined the epidemiological changes in Plasmodium species distribution in Malaysia. Malaria cases shifted from predominantly caused by human malaria to simian malaria, which accounted for the majority of indigenous cases particularly in Malaysia Borneo. Therefore, malaria case notification and prompt malaria diagnosis in regions where health services are limited in Malaysia should be strengthened and reinforced to achieving the final goal of malaria elimination in the country.

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Evaluation of new multiplex PCR primers for the identification ofPlasmodium species found in Sabah, Malaysia

Cited by 15 publications

References 40 publications

Plasmodium malariae in the Colombian Amazon region: you don’t diagnose what you don’t suspect

Plasmodium malariae in the Colombian Amazon region: you don’t diagnose what you don’t suspect

Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia

Malaria distribution and performance of malaria diagnostic methods in Malaysia (1980-2019): a systematic review

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