High-Throughput Multiplex Quantitative Polymerase Chain Reaction Method for Giardia lamblia and Cryptosporidium Species Detection in Stool Samples

Nurminen, Noora; Juuti, Rosa; Oikarinen, Sami; Fan, Yue‐Mei; Lehto, Kirsi‐Maarit; Mangani, Charles; Maleta, Kenneth; Ashorn, Per; Hyöty, Heikki

doi:10.4269/ajtmh.15-0054

Cited by 10 publications

(9 citation statements)

References 21 publications

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“…The Cryptosporidium spp. prevalence in the Malawian samples in the present study was 20%, which is approximately two to four times higher than the prevalence (5.9 to 11%) reported in three other studies from the same region [28–30]. The majority of Cryptosporidium isolates from Malawi as well as Kenya were typed as C. hominis and in line with previous observations [9, 27, 30].…”

Section: Discussionsupporting

confidence: 90%

Performance of three rapid diagnostic tests for the detection of Cryptosporidium spp. and Giardia duodenalis in children with severe acute malnutrition and diarrhoea

et al. 2019

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BackgroundThere is significant need for accurate diagnostic tools for Cryptosporidium spp. and Giardia duodenalis infections in resource limited countries where diarrhoeal disease caused by these parasites is often prevalent. The present study assessed the diagnostic performance of three commercially available rapid diagnostic tests (RDTs) based on faecal-antigen detection for Cryptosporidium spp. and/or G. duodenalis infections in stool samples of children admitted with severe acute malnutrition (SAM) and diarrhoea. An established multiplex PCR was used as reference test.MethodsStool samples from children with SAM and diarrhoea enrolled in a randomized controlled trial (registered at clinicaltrials.gov/ct2/show/NCT02246296) in Malawi (n = 175) and Kenya (n = 120) between December 2014 and December 2015 were analysed by a multiplex PCR for the presence of Cryptosporidium spp., G. duodenalis or Entamoeba histolytica parasite DNA. Cryptosporidium-positive samples were species typed using restriction fragment length polymorphism analysis. A sub-sample of the stool specimens (n = 236) was used for testing with three different RDTs. Diagnostic accuracy of the tests under evaluation was assessed using the results of PCR as reference standard using MedCalc software. Pearson Chi-square test and Fisher’s exact test were used to determine (significant) difference between the number of cryptosporidiosis or giardiasis cases found by PCR in Malawi and Kenya. The overall diagnostic accuracy of each RDT was calculated by plotting a receiver operating characteristic (ROC) curve for each test and to determine the area under the curve (AUC) using SPSS8 software.ResultsPrevalence of Cryptosporidium spp. by PCR was 20.0 and 21.7% in Malawi and Kenya respectively, mostly C. hominis. G. duodenalis prevalence was 23.4 and 5.8% in Malawi and Kenya respectively. E. histolytica was not detected by PCR. RDT testing followed the same pattern of prevalence. RDT sensitivities ranged for cryptosporidiosis from 42.9 to 76.9% and for G. duodenalis from 48.2 to 85.7%. RDT specificities ranged from 88.4 to 100% for Cryptosporidium spp. and from 91.2 to 99.2% for G. duodenalis infections. Based on the estimated area under the curve (AUC) values, all tests under evaluation had an acceptable overall diagnostic accuracy (> 0.7), with the exception of one RDT for Cryptosporidium spp. in Malawi.ConclusionsAll three RDTs for Cryptosporidium spp. and Giardia duodenalis evaluated in this study have a moderate sensitivity, but sufficient specificity. The main value of the RDTs is within their rapidness and their usefulness as screening assays in surveys for diarrhoea.

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Section: Discussionsupporting

confidence: 90%

Performance of three rapid diagnostic tests for the detection of Cryptosporidium spp. and Giardia duodenalis in children with severe acute malnutrition and diarrhoea

et al. 2019

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“…Nucleic acids were isolated using QIAamp Viral RNA Mini Kit (Qiagen, Hilden, Germany) from the 10% faecal suspension and real-time PCR was used to detect enteroviruses, rhinoviruses, noroviruses, parechoviruses and rotaviruses [21, 22] as well as Giardia lamblia and Cryptosporidium (spp.) parasites as previously reported [23]. The primers and probes used for PCR were shown in supplementary Table S1.…”

Section: Methodsmentioning

confidence: 99%

High prevalence of selected viruses and parasites and their predictors in Malawian children

et al. 2019

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Enteric pathogens have been related to child undernutrition. Whereas there are lots of data on enteric bacterial microbiota and infections, much less is known about the incidence of prevalence of intestinal colonisation with viruses or important parasitic species. This study assessed the presence of selected viruses and parasites in stools of 469, 354, 468 Malawian children at 6, 12 and 18 months. We also assessed environmental predictors of the presence of viruses and parasites among 6-month infants. Microbial presence was documented using real-time polymerase chain reaction (PCR). Enteroviruses were identified in 68%, 80% and 81% of the stool samples at 6, 12 and 18 months children, rhinovirus in 28%, 18% and 31%, norovirus in 24%, 22% and 16%, parechovirus in 23%, 17% and 17%, rotavirus in 3%, 1% and 0.6%, Giardia lamblia in 9.6%, 23.5% and 26%, and Cryptosporidium (spp.) in 6%, 8% and 2% of the 6, 12 and 18 months stool samples. Dry season (May–October) was associated with a low infection rate of enterovirus, norovirus and Cryptosporidium (spp.). Higher father's education level, less number of person in the household and higher sanitation were associated with a low infection rate of enterovirus, norovirus and rotavirus, respectively. The results suggest that the prevalence of asymptomatic viral and parasitic infections is high among Malawian children and that the family's living conditions and seasonality influence the rate of infections.

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“…We used real‐time PCR to characterise enterovirus, rotavirus, norovirus, parechovirus and rhinovirus , and parasites G . lamblia and Cryptosporidium species . We considered a specimen as a positive sample if two or three of the triplicate PCR runs gave a positive test result.…”

Section: Methodsmentioning

confidence: 99%

Presence of Giardia lamblia in stools of six‐ to 18‐month old asymptomatic Malawians is associated with children's growth failure

et al. 2019

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Aim Despite high pathogen burden and malnutrition in low‐income settings, knowledge on relationship between asymptomatic viral or parasitic infections, nutrition and growth is insufficient. We studied these relationships in a cohort of six‐month‐old Malawian infants. Methods As part of a nutrient supplementation trial for 12 months, we documented disease symptoms of 840 participant daily and anthropometric measurements every three months. Stool specimens were collected every six months and analysed for Giardia lamblia, Cryptosporidium species and enterovirus, rotavirus, norovirus, parechovirus and rhinovirus using polymerase chain reaction (PCR). The prevalence of the microbes was compared to the children's linear growth and the dietary. Results The prevalence of the microbes was similar in every intervention group. All age groups combined, children negative for G. lamblia had a mean standard deviation (SD) of −0.01 (0.49) change in length‐for‐age Z‐score (LAZ), compared to −0.12 (0.045) among G. lamblia positive children (difference −0.10, 95% CI −0.21 to −0.00, p = 0.047). The LAZ change difference was also statistically significant (p = 0.042) at age of 18–21 months but not at the other time points. Conclusion Asymptomatic G. lamblia infection was mainly associated with growth reduction in certain three‐month periods. The result refers to the chronic nature of G. lamblia infection.

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High-Throughput Multiplex Quantitative Polymerase Chain Reaction Method for Giardia lamblia and Cryptosporidium Species Detection in Stool Samples

Cited by 10 publications

References 21 publications

Performance of three rapid diagnostic tests for the detection of Cryptosporidium spp. and Giardia duodenalis in children with severe acute malnutrition and diarrhoea

Performance of three rapid diagnostic tests for the detection of Cryptosporidium spp. and Giardia duodenalis in children with severe acute malnutrition and diarrhoea

High prevalence of selected viruses and parasites and their predictors in Malawian children

Presence of Giardia lamblia in stools of six‐ to 18‐month old asymptomatic Malawians is associated with children's growth failure

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