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Fulltext Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis

Ngui R, Lim YA, Chua KH

PLoS One, 2012;7(7):e41996.
PMID: 22844538 DOI: 10.1371/journal.pone.0041996

Hookworm infections are still endemic in low and middle income tropical countries with greater impact on the socioeconomic and public health of the bottom billion of the world's poorest people. In this study, a real-time polymerase chain reaction (PCR) coupled with high resolution melting-curve (HRM) analysis was evaluated for an accurate, rapid and sensitive tool for species identification focusing on the five human hookworm species.
Spatially varying correlation between environmental conditions and human leptospirosis in Sarawak, Malaysia

Kira R, Bilung LM, Ngui R, Apun K, Su'ut L

Trop Biomed, 2021 Jun 01;38(2):31-39.
PMID: 33973570 DOI: 10.47665/tb.38.2.034

The spatial distribution of environmental conditions may influence the dynamics of vectorborne diseases like leptospirosis. This study aims to investigate the global and localised relationships between leptospirosis with selected environmental variables. The association between environmental variables and the spatial density of geocoded leptospirosis cases was determined using global Poisson regression (GPR) and geographically weighted Poisson regression (GWPR). A higher prevalence of leptospirosis was detected in areas with higher water vapour pressure (exp(â): 1.12; 95% CI: 1.02 - 1.25) and annual precipitation (exp(â): 1.15; 95% CI: 1.02 - 1.31), with lower precipitation in the driest month (exp(â): 0.85; 95% CI: 0.75 - 0.96) and the wettest quarter (exp(â): 0.88; 95% CI: 0.77 - 1.00). Water vapor pressure (WVP) varied the most in the hotspot regions with a standard deviation of 0.62 (LQ: 0.15; UQ; 0.99) while the least variation was observed in annual precipitation (ANNP) with a standard deviation of 0.14 (LQ: 0.11; UQ; 0.30). The reduction in AICc value from 519.73 to 443.49 indicates that the GWPR model is able to identify the spatially varying correlation between leptospirosis and selected environmental variables. The results of the localised relationships in this study could be used to formulate spatially targeted interventions. This would be particularly useful in localities with a strong environmental or socio-demographical determinants for the transmission of leptospirosis.
A Case of Zoonotic Ancylostoma ceylanicum Infection in a Suburban Area of Selangor, Malaysia

Heo CC, Rafiz AR, Ngui R

Acta Parasitol, 2021 Oct 06.
PMID: 34617193 DOI: 10.1007/s11686-021-00478-1

INTRODUCTION: The animal hookworm, Ancylostoma ceylanicum, is a dominant hookworm species of dogs and cats. However, it has increasingly been found infecting humans in Southeast Asia.
PURPOSE: We report an autochthonous case of A. ceylanicum in a suburban area of Selangor, Malaysia. A 66-year-old Indian lady who is an avid gardener presented with chronic diarrhea of 4 months' duration.
METHODS: The patient was examined clinically and colonoscopy was performed. Adult parasites obtained via colonoscopy were subjected to microscopy and molecular investigations.
RESULTS: Clinical examinations were unremarkable, and blood investigation revealed normochromic normocytic anemia. Stool occult blood was positive but negative for ova, cyst and adult parasites. Colonoscopy performed showed multiple diverticulae and worm infestation from the terminal ileum to sigmoid colon. Morphological examination on the adult worms showed the specific characteristics of Ancylostoma species. Molecular investigations further confirmed the nematode as Ancylostoma ceylanicum. She was treated with albendazole 400 mg daily for 3 days with symptomatic improvements sustained 3 months later. It is suspected that the patient had ingested or contacted soil contaminated with filariform larvae while gardening.
CONCLUSION: Information on the A. ceylanicum infection in humans, especially in urban and suburban areas, is limited, necessitating further epidemiological and clinical studies.
Serological and molecular detection of Strongyloides stercoralis infection among an Orang Asli community in Malaysia

Ahmad AF, Hadip F, Ngui R, Lim YA, Mahmud R

Parasitol Res, 2013 Aug;112(8):2811-6.
PMID: 23666229 DOI: 10.1007/s00436-013-3450-z

Detection of Strongyloides stercoralis infection particularly in asymptomatic individuals is often hampered due to the lack of standard diagnostic tools. In this study, the use of serological and molecular approaches were investigated for the detection of S. stercoralis infection among an Orang Asli (indigenous) community following a preliminary detection by microscopic examination of faecal samples. Out of 54 individuals studied, 17/54 (31.5%) were detected to be positive for S. stercoralis infection by enzyme-linked immunosorbent assay (ELISA), compared to 0/54 (0%) by faecal examination. Further confirmation performed by a nested polymerase chain reaction (PCR) using DNA extracted from faecal samples of these 17 individuals yielded 3/17 (17.6%) positives for S. stercoralis DNA amplification. No amplification was seen with the other 37 faecal samples, which were negative by microscopy and ELISA. As the high ELISA positive results were suspected to be false-positives, ELISA is not recommended for use as a detection tool but may be beneficial for evaluating the effectiveness of anti-Strongyloides drugs. The present finding indicated that PCR should be considered as an alternative diagnostic tool for the detection of S. stercoralis infection.
Fulltext Prevalence and risk factors of intestinal parasitism in rural and remote West Malaysia

Ngui R, Ishak S, Chuen CS, Mahmud R, Lim YA

PLoS Negl Trop Dis, 2011;5(3):e974.
PMID: 21390157 DOI: 10.1371/journal.pntd.0000974

Intestinal parasitic infections (IPIs) have a worldwide distribution and have been identified as one of the most significant causes of illnesses and diseases among the disadvantaged population. In Malaysia, IPIs still persist in some rural areas, and this study was conducted to determine the current epidemiological status and to identify risk factors associated with IPIs among communities residing in rural and remote areas of West Malaysia.
Intestinal parasites in various animals at a zoo in Malaysia

Lim YA, Ngui R, Shukri J, Rohela M, Mat Naim HR

Vet Parasitol, 2008 Oct 20;157(1-2):154-9.
PMID: 18723289 DOI: 10.1016/j.vetpar.2008.07.015

A survey was undertaken to investigate the prevalence of intestinal parasites from different groups of mammals housed in a zoological garden in Malaysia. A total of 197 faecal samples were collected randomly from various primates (99), hoofed mammals (70) and feline (28). It was discovered that 89.3% of feline, 54.5% of primates and 45.7% of hoofed mammals were infected with intestinal parasites. Intestinal parasites found in primates were Balantidium coli (19.2%), Cryptosporidium spp. (14.1%), hookworm (10.1%), Trichuris spp. (5.1%), Ascaris (4.0%) and Blastocystis spp. (2.0%). For hoofed mammals, hookworm had the highest prevalence (34.3%) followed by Trichuris spp. and Cryptosporidium spp. (5.7%). Meanwhile, for feline, Toxocara cati was the most prevalent (64.3%), followed by Cryptosporidium spp. (14.3%), Spirometra spp. (7.1%), and hookworm (3.6%). Animals that were infected were all asymptomatic with low parasite load. Routine monitoring of the presence of parasites in animals kept in the zoo is imperative in assisting zoo management in the formulation and implementation of preventive and control measures against the spread of infectious parasitic diseases among animals within the zoo or to humans.
Fulltext Current status of parasitic infections among Pangkor Island community in Peninsular Malaysia

Ahmad AF, Ngui R, Muhammad Aidil R, Lim YA, Rohela M

Trop Biomed, 2014 Dec;31(4):836-43.
PMID: 25776610 MyJurnal

Limited data is available on the prevalence of parasitic infections among the island communities in Malaysia with most studies performed between 1960s-1980s. This study was conducted to determine the current prevalence status of parasitic infections among communities living in Pangkor Island Peninsular Malaysia. A total of 131 stool and 298 serum samples were collected and subjected to microscopic examination for intestinal protozoa and helminths and detection of Toxoplasma gondii antibodies using commercial ELISA kits respectively. In addition, thin and thick peripheral blood films were microscopically screened for the presence of Plasmodium spp. and microfilariae respectively. The overall prevalence of intestinal parasitic infections among Pangkor Island community was 9.9% (13/131) with T. trichiura (5.3%) being the most common intestinal parasite detected. Toxoplasmosis was reported in almost 60% of the community with the seroprevalence being significantly high among females (64.7%) compared to males (52.8%) (p=0.038). None of those examined samples were infected with intestinal sarcocystosis, malaria and filariasis. This study revealed that the prevalence of intestinal parasitic infections among Pangkor Island community has been greatly reduced compared to that reported 35 years ago. Massive improvements in the socioeconomic status, personal hygiene, water facilities and sanitation may have contributed to the low prevalence of parasitic infections in this community. Nevertheless, further studies still need to be performed to determine the possible risk factors for the high prevalence of toxoplasmosis in this community.
Tongue necrosis secondary to mucormycosis in a diabetic patient: A first case report in Malaysia

Fattah SY, Hariri F, Ngui R, Husman SI

J Mycol Med, 2018 Sep;28(3):519-522.
PMID: 30205883 DOI: 10.1016/j.mycmed.2018.06.004

Mucormycosis is a rare fungal infection and high mortality that commonly affects patients with the weakened immune system. We present an unusual case of tongue necrosis probably due to the healthcare-associated mucormycosis (HCM) in a diabetic patient. Although cannot be proved with certainty, we surmise that intubation as a risk factor in our case. The diagnosis was confirmed by histopathological examination (HPE) of the necrotic tissue specimen. The patient was responded well to lipid complex amphotericin B (250mg) regime after surgery. Subsequent follow up revealed that no signs of recurrence. Early, recognition, diagnosis, prompt treatment and awareness among clinician are representing the most effective way of managing the disease.
Fulltext Evaluation of sticky traps for adult Aedes mosquitoes in Malaysia: a potential monitoring and surveillance tool for the efficacy of control strategies

Roslan MA, Ngui R, Vythilingam I, Sulaiman WYW

J Vector Ecol, 2017 12;42(2):298-307.
PMID: 29125255 DOI: 10.1111/jvec.12270

The present study compared the performance of sticky traps in order to identify the most effective and practical trap for capturing Aedes aegypti and Aedes albopictus mosquitoes. Three phases were conducted in the study, with Phase 1 evaluating the five prototypes (Models A, B, C, D, and E) of sticky trap release-and-recapture using two groups of mosquito release numbers (five and 50) that were released in each replicate. Similarly, Phase 2 compared the performance between Model E and the classical ovitrap that had been modified (sticky ovitrap), using five and 50 mosquito release numbers. Further assessment of both traps was carried out in Phase 3, in which both traps were installed in nine sampling grids. Results from Phase 1 showed that Model E was the trap that recaptured higher numbers of mosquitoes when compared to Models A, B, C, and D. Further assessment between Model E and the modified sticky ovitrap (known as Model F) found that Model F outperformed Model E in both Phases 2 and 3. Thus, Model F was selected as the most effective and practical sticky trap, which could serve as an alternative tool for monitoring and controlling dengue vectors in Malaysia.
Fulltext Community surveillance of Aedes albopictus associated with Wolbachia detection in low-rise residential areas in Selangor, Malaysia

Roslan MA, Ngui R, Vythilingam I, Wan Sulaiman WY

J Vector Ecol, 2022 Dec;47(2):142-152.
PMID: 36314668 DOI: 10.52707/1081-1710-47.2.142

The study assessed the distribution of Malaysian Ae. albopictus adults associated with Wolbachia detection in low-rise residential areas using a modified sticky ovitrap (MSO). The relationship between Ae. albopictus and climatological parameters were also determined. Fifty-two weeks of surveillance using 273 MSOs were conducted in four installation areas of eleven sampling sites. Specimens were subjected to PCR using wsp-specific primers for Wolbachia detection. The relationship between climatological parameters and Ae. albopictus captured were analyzed using Spearman rank correlation coefficient test. The majority of Ae. albopictus were captured in residential houses (87%), followed by playgrounds or parks (11.5%), guardhouses (1%), and community halls (0.5%). Most of the specimens (92%) were superinfected with wAlbA and wAlbB strains. A positive correlation with no significant association was found for rainfall (r = 0.015, P = 0.072), relative humidity (r = 0.005, P = 0.526), minimum temperature (r = 0.005, P = 0.516), and mean temperature (r = 0.003, P = 0.689). MSO effectively captured a high number of Ae. albopictus that was determined to be the predominant mosquito species found in low-rise residential areas. The adult collection is not only influenced by climatological parameters but also by other factors, including environmental conditions and general sanitation status.
Fulltext Expression of Ascaris lumbricoides putative virulence-associated genes when infecting a human host

Mohd-Shaharuddin N, Lim YAL, Ngui R, Nathan S

Parasit Vectors, 2021 Mar 23;14(1):176.
PMID: 33757548 DOI: 10.1186/s13071-021-04680-y

BACKGROUND: Ascaris lumbricoides is the most common causative agent of soil-transmitted helminth infections worldwide, with an estimated 450 million people infected with this nematode globally. It is suggested that helminths are capable of evading and manipulating the host immune system through the release of a spectrum of worm proteins which underpins their long-term survival in the host. We hypothesise that the worm overexpresses these proteins when infecting adults compared to children to cirvumvent the more robust defence mechanisms of adults. However, little is known about the parasite's genes and encoded proteins involved during A. lumbricoides infection. Hence, this study was conducted to assess the expression profile of putative virulence-associated genes during an active infection of adults and children.
METHODS: In this study, quantitative PCR was performed to evaluate the expression profile of putative virulence-associated genes in A. lumbricoides isolated from infected children and adults. The study was initiated by collecting adult worms expelled from adults and children following anthelminthic treatment. High-quality RNA was successfully extracted from each of six adult worms expelled by three adults and three children, respectively. Eleven putative homologues of helminth virulence-associated genes reported in previous studies were selected, primers were designed and specific amplicons of A. lumbricoides genes were noted. The expression profiles of these putative virulence-associated genes in A. lumbricoides from infected adults were compared to those in A. lumbricoides from infected children.
RESULTS: The putative virulence-associated genes VENOM, CADHERIN and PEBP were significantly upregulated at 166-fold, 13-fold and fivefold, respectively, in adults compared to children. Conversely, the transcription of ABA-1 (fourfold), CATH-L (threefold) and INTEGRIN (twofold) was significantly suppressed in A. lumbricoides from infected adults.
CONCLUSIONS: On the basis of the expression profile of the putative virulence-associated genes, we propose that the encoded proteins have potential roles in evasion mechanisms, which could guide the development of therapeutic interventions.
Fulltext Epidemiological and genetic data supporting the transmission of Ancylostoma ceylanicum among human and domestic animals

Ngui R, Lim YA, Traub R, Mahmud R, Mistam MS

PLoS Negl Trop Dis, 2012;6(2):e1522.
PMID: 22347515 DOI: 10.1371/journal.pntd.0001522

Currently, information on species-specific hookworm infection is unavailable in Malaysia and is restricted worldwide due to limited application of molecular diagnostic tools. Given the importance of accurate identification of hookworms, this study was conducted as part of an ongoing molecular epidemiological investigation aimed at providing the first documented data on species-specific hookworm infection, associated risk factors and the role of domestic animals as reservoirs for hookworm infections in endemic communities of Malaysia.
Fulltext Antibody prevalence and factors associated with exposure to Orientia tsutsugamushi in different aboriginal subgroups in West Malaysia

Tay ST, Mohamed Zan HA, Lim YA, Ngui R

PLoS Negl Trop Dis, 2013;7(8):e2341.
PMID: 23936576 DOI: 10.1371/journal.pntd.0002341

BACKGROUND: Limited data is available on the current status of scrub typhus infection in the aboriginal population in Malaysia. This study was aimed to provide recent data on the degree of exposure of 280 individuals from seven aboriginal subgroups to Orientia tsutsugamushi (causative agent of scrub typhus) in West Malaysia. The environment, socioeconomic and behavioural risk factors associated with the disease were also investigated.
METHODS/FINDINGS: The antibody prevalence to O. tsutsugamushi ranged from 0 to 36.4% in seven subgroups, with high prevalence rates noted in subgroups involved in agricultural activity and the lowest prevalence rates noted in subgroups whose main occupations were associated to fishing. Univariate analysis indicated populations with age above 18 years (OR = 1.15, 95% CI = 1.02-1.30, P = 0.015), working (OR = 1.99, 95% CI = 1.01-3.92, P = 0.044), working at agriculture area (OR = 1.18, 95% CI = 0.98-1.42, P = 0.031), receiving household income less than US$ 166.7 (RM500) per month (OR = 2.43, 95% CI = 1.16-5.11, P = 0.016) and having close contact with animal pets (OR = 4.06, 95% CI = 1.20-13.76, P = 0.016) are significantly associated with exposure to O. tsutsugamushi. Multivariate analysis confirms that participants who are above 18 years old, receiving household income less than US$ 166.7 (RM500) per month and having close contact with animal pets are 3.6 times (95% CI = 1.81-7.03, P<0.001), 1.3 times (95% CI = 1.14-1.64, P = 0.002) and 1.2 times (95% CI = 1.05-1.06, P = 0.006) more likely to have exposure to O. tsutsugamushi, respectively.
CONCLUSION: The present study indicates that scrub typhus is still an important disease in the aboriginal population in Malaysia. Awareness about the disease and education on the preventive measures are important in reducing the risk of acquiring scrub typhus in the population studied.
Fulltext Neglected tropical diseases among two indigenous subtribes in peninsular Malaysia: highlighting differences and co-infection of helminthiasis and sarcocystosis

Lee SC, Ngui R, Tan TK, Muhammad Aidil R, Lim YA

PLoS One, 2014;9(9):e107980.
PMID: 25248116 DOI: 10.1371/journal.pone.0107980

Soil-transmitted helminth (STH) infections have been documented among these minority groups since 1938. However the prevalence of STH is still high among these communities. Most studies tend to consider the Orang Asli (indigenous) as a homogenous group. In contrary, different subtribes have their own cultural practices. To understand this variation better, we studied the prevalence and associated factors of STH and other gut parasitic infections among two common subtribes (i.e. Temuan and Temiar). Results showed that the prevalence of the overall STH infections was higher in the Temuan subtribe (53.2% of 171) compared to the Temiar subtribe (52.7% of 98). Trichuris trichiura (46.2%) was the most prevalent parasite in the Temuan subtribe, followed by Ascaris spp. (25.7%) and hookworm (4.1%). In contrast, Ascaris spp. (39.8%) was more prevalent among the Temiar subtribe, preceded by T. trichiura (35.7%) and finally hookworm (8.3%). There were also co-infections of helminthiasis and intestinal protozoa among both Temuan and Temiar subtribes with rates being three times higher among the Temiar compared to Temuan. The most common co-infection was with Entamoeba histolytica/dispar/moshkovskii (n = 24; 24.5%, 16.0-33.0), followed by Giardia spp. (n = 3; 3.1%, -0.3-6.5). In Temuan, STH infection individuals were also infected with Entamoeba histolytica/dispar/moshkovskii (n = 11; 6.4%, 5.0-13.8), Cryptosporidium spp. (n = 3, 1.8%, -0.2-3.8) and Giardia spp. (n = 2, 1.2%, -0.4-2.8). In comparison, there was no Cryptosporidium spp. detected among the Temiar. However, it was interesting to note that there was an occurrence of co-infection of intestinal helminthiasis and sarcocystosis (intestinal) in a Temiar individual. The last report of sarcocystosis (muscular) among the Orang Asli was in 1978. The present study highlighted the importance of understanding the variation of infections amongst the different Orang Asli subtribes. It is vital to note these differences and use this knowledge to customise effective control measures for the various subtribes.
Spatial density of Aedes distribution in urban areas: a case study of breteau index in Kuala Lumpur, Malaysia

Aziz S, Aidil RM, Nisfariza MN, Ngui R, Lim YA, Yusoff WS, et al.

J Vector Borne Dis, 2014 Jun;51(2):91-6.
PMID: 24947215

Dengue fever (DF) is a major vector-borne disease in Malaysia. The incidences of DF in Malaysia are caused by viruses transmitted through the bites of infected female Aedes albopictus and Ae. aegypti mosquitoes. This study aims to establish the spatial density of mosquito population or breteau index (BI) in the areas of Kuala Lumpur using geographic information system (GIS), remote sensing (RS) and spatial statistical tools.
Fulltext Zoonotic Ancylostoma ceylanicum infection detected by endoscopy

Ngui R, Lim YA, Ismail WH, Lim KN, Mahmud R

Am J Trop Med Hyg, 2014 Jul;91(1):86-8.
PMID: 24891471 DOI: 10.4269/ajtmh.13-0756

We report a case of Ancylostoma ceylanicum infection detected by endoscopy. It was diagnosed and confirmed using polymerase chain reaction (PCR) and DNA sequencing. The patient is a 58-year-old Malaysian woman who lives in a rural area, where uncontrolled populations of stray and semidomesticated dogs live in close proximity with humans.
Fulltext Co-infection of Haemonchus contortus and Trichostrongylus spp. among livestock in Malaysia as revealed by amplification and sequencing of the internal transcribed spacer II DNA region

Tan TK, Panchadcharam C, Low VL, Lee SC, Ngui R, Sharma RS, et al.

BMC Vet Res, 2014;10:38.
PMID: 24502557 DOI: 10.1186/1746-6148-10-38

Haemonchus contortus and Trichostrongylus spp. are reported to be the most prevalent and highly pathogenic parasites in livestock, particularly in small ruminants. However, the routine conventional tool used in Malaysia could not differentiate the species accurately and therefore limiting the understanding of the co-infections between these two genera among livestock in Malaysia. This study is the first attempt to identify the strongylids of veterinary importance in Malaysia (i.e., H. contortus and Trichostrongylus spp.) by amplification and sequencing of the Internal Transcribed Spacer II DNA region.
Genetic characterization of the partial mitochondrial cytochrome oxidase c subunit I (cox 1) gene of the zoonotic parasitic nematode, Ancylostoma ceylanicum from humans, dogs and cats

Ngui R, Mahdy MA, Chua KH, Traub R, Lim YA

Acta Trop, 2013 Oct;128(1):154-7.
PMID: 23774318 DOI: 10.1016/j.actatropica.2013.06.003

Ancylostoma ceylanicum is the only zoonotic hookworm species that is able to produce patent infections in humans with the majority of cases reported in South East Asia. Over the past few years, there have been an increasing number of studies investigating the prevalence of this parasitic zoonosis using molecular diagnostic tools and a single genetic locus as marker for species identification. As there can be limitations in using a single genetic locus for epidemiological studies and genetic discrimination, the complementary use of a more variable locus will provide additional evidence to support the zoonotic exchange of hookworm species between humans and animals. In the present study, the cytochrome c oxidase subunit 1 (cox 1) sequence of A. ceylanicum from positive human and animal fecal samples were determined and compared with published reference sequences. Phylogenetic analysis demonstrated that isolates of A. ceylanicum were divided into two clusters, one consisting 3 human isolates and the other comprising 19 isolates of human and animal origin from different geographical locations within Malaysia. The two groups of A. ceylanicum could be distinguished from one another through five fixed nucleotide differences at locations 891, 966, 1008, 1077 and 1083. The detection of genetically distinct groups and considerable level of genetic variation within the cox 1 sequence of A. ceylanicum might suggest potential haplotype-linked differences in zoonotic, epidemiological and pathobiological characteristics, a hypothesis that still needs further investigation.
Fulltext Molecular identification of human hookworm infections in economically disadvantaged communities in Peninsular Malaysia

Ngui R, Ching LS, Kai TT, Roslan MA, Lim YA

Am J Trop Med Hyg, 2012 May;86(5):837-42.
PMID: 22556084 DOI: 10.4269/ajtmh.2012.11-0446

Species identification of human hookworm infections among eight communities in rural areas of Peninsular Malaysia was determined during 2009-2011. Fecal samples were examined by microscopy and subsequently, the internal transcribed spacer 2 and 28S ribosomal RNA region of Necator americanus and Ancylostoma spp. were sequenced. Overall, 9.1% (58 of 634) were identified positive by microscopy for hookworm infection, and 47 (81.0%) of 58 were successfully amplified and sequenced. Sequence comparison found that N. americanus (87.2%) was the most predominant hookworm identified, followed by Ancylostoma ceylanicum (23.4%). No A. duodenale infection was detected in this study. Detection of A. ceylanicum in humans highlighted the zoonotic transmission among humans living near dogs. Thus, implementation of effective control measures for hookworm infections in future should seriously consider this zoonotic implication.
Differentiating Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii using nested polymerase chain reaction (PCR) in rural communities in Malaysia

Ngui R, Angal L, Fakhrurrazi SA, Lian YL, Ling LY, Ibrahim J, et al.

Parasit Vectors, 2012;5:187.
PMID: 22947430

In this study, a total of 426 human faecal samples were examined for the presence of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii infection via a combination of microscopic examination and nested polymerase chain reaction (PCR) targeting 16S ribosomal RNA of Entamoeba species.

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