Improved diagnosis of <i>Trichuris trichiura</i> by using a bead-beating procedure on ethanol preserved stool samples prior to DNA isolation and the performance of multiplex real-time PCR for intestinal parasites

Kaisar, Maria M. M.; Brienen, Eric A. T.; Djuardi, Yenny; Sartono, Erliyani; Yazdanbakhsh, Maria; Verweij, Jaco J.; Supali, Taniawati; Lieshout, Lisette van

doi:10.1017/s0031182017000129

Cited by 73 publications

(99 citation statements)

References 38 publications

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“…Interestingly, some authors also reported poorer DNA extraction after this step, depending of the extraction technique used and on the type of beads. For example, in the study of Kaisar et al [ 11 ], mechanical lysis diminished the number of positive samples for G. intestinalis and D. fragilis . In the same way, a multicentric study showed highly variable effects of the bead-beating step on the extraction of Cryptosporidium DNA, especially with the MagNA Lyser ® Green Beads which impacted it negatively [ 16 ].…”

Section: Discussionmentioning

confidence: 99%

Comparison of three commercial multiplex PCR assays for the diagnosis of intestinal protozoa

et al. 2018

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Although microscopic examination of stool samples remains the reference method for the diagnosis of intestinal protozoal infections, these techniques are time-consuming and require operators who are experienced and well trained. Molecular biology seems to offer performances at least equivalent in terms of sensitivity and specificity for certain parasites. This study aimed to compare three multiplex PCR assays on 93 prospectively collected positive stools (prospective cohort) and a panel of 12 more Cryptosporidium-positive samples (Cryptosporidium panel). On the prospective cohort, the sensitivity was 89%, 64% and 41% for Giardia sp. detection for BD MaxTM, G-DiaParaTM and RIDA®GENE, respectively and 75%, 100% and 100% for C. parvum/hominis detection. The sensitivity of the RIDA®GENE assay for all Cryptosporidium species was 100%, and for D. fragilis 71%. All the techniques obtained the same results for E. histolytica detection, with one positive sample. All species in the Cryptosporidium panel were identified by the RIDA®GENE PCR. The BD MaxTM and G-DiaParaTM assays detected only C. parvum/hominis with the exception of one positive sample for C. meleagridis. No assay showed satisfactory results for all parasites simultaneously, and the DNA extraction seems to be the critical step. More studies are needed to standardize this procedure.

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Section: Discussionmentioning

confidence: 99%

Comparison of three commercial multiplex PCR assays for the diagnosis of intestinal protozoa

et al. 2018

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“…The POL method is similar to the technique of helminth DNA extraction described by Verweij (35) except the missing bead mill procedure, which was not available in our laboratories at the time of the study. The POL method was the only alternative method (with good results for protozoans) known, which could be used without a cell lysing instrument (9,18,22,36).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Two DNA Extraction Methods for Detection of Strongyloides stercoralis Infection

et al. 2018

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is present worldwide, but its prevalence is still uncertain, mainly due to the lack of sensitivity of diagnostic methods. Molecular techniques are under development, but a standardized protocol is still unavailable. We compared the sensitivity of real-time PCR, using two extraction protocols, with that of the Baermann technique. Samples were collected in the framework of the baseline screening of a randomized clinical trial evaluating moxidectin against in Lao People's Democratic Republic. Two stool samples from each participant were processed by the Baermann method, and one subsample was processed by PCR. DNA was extracted using the QIAamp DNA stool minikit based on the standard protocol for the QIAamp DNA minikit (QIA) and using a modification of the QIA procedure (POL). Subsequently, all extracted samples were analyzed by real-time PCR. Overall, 95 samples were analyzed by the three diagnostic methods. Sixty-nine (72.6%) samples were positive according to the Baermann method, 25 (26.3%) by the QIA method, and 62 (65.3%) by the POL method. The sensitivities were 86% (95% confidence interval [CI], 76.7 to 92.9), 31.0% (95% CI, 21.3 to 42.6), and 78.0% (95% CI, 66.8 to 86.1) for the Baermann, QIA, and POL methods, respectively. The sensitivities calculated for each day of the Baermann method separately were 60% (48.4 to 70.8%) and 64% (52.2 to 74.2%) for days 1 and 2, respectively. In conclusion, the POL method revealed a good performance and was comparable to the Baermann test performed on two stool samples and superior to the Baermann method performed on one stool sample. Additional studies are needed to standardize a PCR protocol for diagnosis.

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“…Two panels of multiplex real‐time PCR were used to detect and quantify soil transmitted helminths and intestinal protozoa. Panel 1 targets hookworm ( Ancylostoma duodenale, Necator americanus ), Ascaris lumbricoides , Trichuris trichuria and Strongyloides stercoralis , while panel 2 targets Entamoeba histolytica, Dientamoeba fragilis, Giardia lamblia and Cryptosporidium spp. .…”

Section: Methodsmentioning

confidence: 99%

BCG scar, socioeconomic and nutritional status: a study of newborns in urban area of Makassar, Indonesia

Amaruddin

Wahyuni

Hamid

et al. 2019

Tropical Med Int Health

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SummaryObjectiveTo investigate factors that determine the response to Bacille Calmette–Guérin (BCG) vaccination in urban environments with respect to socioeconomic status (SES), prenatal exposure to infections or newborn's nutritional status.MethodsThe study was conducted in an urban area, in Makassar, Indonesia. At baseline, 100 mother and newborns pair from high and low SES communities were included. Intestinal protozoa, soil transmitted helminths, total IgE, anti‐Hepatitis A Virus IgG and anti‐Toxoplasma IgG were measured to determine exposure to infections. Information on gestational age, birth weight/height and delivery status were collected. Weight‐for‐length z‐score, a proxy for newborns adiposity, was calculated. Leptin and adiponectin from cord sera were also measured. At 10 months of age, BCG scar size was measured from 59 infants. Statistical modelling was performed using multiple linear regression.ResultsBoth SES and birth nutritional status shape the response towards BCG vaccination at 10 months of age. Infants born to low SES families have smaller BCG scar size compared to infants born from high SES families and total IgE contributed to the reduced scar size. On the other hand, infants born with better nutritional status were found to have bigger BCG scar size but this association was abolished by leptin levels at birth.ConclusionThis study provides new insights into the importance of SES and leptin levels at birth on the development of BCG scar in 10 months old infants.

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Improved diagnosis of Trichuris trichiura by using a bead-beating procedure on ethanol preserved stool samples prior to DNA isolation and the performance of multiplex real-time PCR for intestinal parasites

Cited by 73 publications

References 38 publications

Comparison of three commercial multiplex PCR assays for the diagnosis of intestinal protozoa

Comparison of three commercial multiplex PCR assays for the diagnosis of intestinal protozoa

Evaluation of Two DNA Extraction Methods for Detection of Strongyloides stercoralis Infection

BCG scar, socioeconomic and nutritional status: a study of newborns in urban area of Makassar, Indonesia

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