Surveillance for soil-transmitted helminths in high-risk county, Mississippi, USA
- Bradbury, Richard, Martin, Lora, Malloch, Lacy, Martin, Maygan, Williams, John, Patterson, Kayla, Sanders, Cameron, Singh, Gurbaksh, Arguello, Irene, Rodriguez, Eduardo, Byers, Paul, Haynie, Lisa, Qvarnstrom, Yvonne, Hobbs, Charlotte
- Authors: Bradbury, Richard , Martin, Lora , Malloch, Lacy , Martin, Maygan , Williams, John , Patterson, Kayla , Sanders, Cameron , Singh, Gurbaksh , Arguello, Irene , Rodriguez, Eduardo , Byers, Paul , Haynie, Lisa , Qvarnstrom, Yvonne , Hobbs, Charlotte
- Date: 2023
- Type: Text , Journal article
- Relation: Emerging Infectious Diseases Vol. 29, no. 12 (2023), p. 2533-2537
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- Description: Recent reports of hookworm infection in Alabama, USA, has prompted surveillance in Mississippi, given the states’ similar environmental conditions. We collected stool specimens from 277 children in Rankin County, Mississippi. Kato–Katz microscopic smear, agar plate culture, and quantitative PCR indicated no soil-transmitted helminths. Nevertheless, further surveillance in other high-risk Mississippi counties is warranted. © 2023 Centers for Disease Control and Prevention (CDC). All rights reserved.
- Authors: Bradbury, Richard , Martin, Lora , Malloch, Lacy , Martin, Maygan , Williams, John , Patterson, Kayla , Sanders, Cameron , Singh, Gurbaksh , Arguello, Irene , Rodriguez, Eduardo , Byers, Paul , Haynie, Lisa , Qvarnstrom, Yvonne , Hobbs, Charlotte
- Date: 2023
- Type: Text , Journal article
- Relation: Emerging Infectious Diseases Vol. 29, no. 12 (2023), p. 2533-2537
- Full Text:
- Reviewed:
- Description: Recent reports of hookworm infection in Alabama, USA, has prompted surveillance in Mississippi, given the states’ similar environmental conditions. We collected stool specimens from 277 children in Rankin County, Mississippi. Kato–Katz microscopic smear, agar plate culture, and quantitative PCR indicated no soil-transmitted helminths. Nevertheless, further surveillance in other high-risk Mississippi counties is warranted. © 2023 Centers for Disease Control and Prevention (CDC). All rights reserved.
Cross-sectional study of soil-transmitted helminthiases in black belt region of Alabama, USA
- Poole, Claudette, Barker, Troy, Bradbury, Richard, Capone, Drew, Chatham, Amy, Handali, Sukwan, Rodriguez, Eduardo, Qvarnstrom, Yvonne, Brown, Joe
- Authors: Poole, Claudette , Barker, Troy , Bradbury, Richard , Capone, Drew , Chatham, Amy , Handali, Sukwan , Rodriguez, Eduardo , Qvarnstrom, Yvonne , Brown, Joe
- Date: 2023
- Type: Text , Journal article
- Relation: Emerging Infectious Diseases Vol. 29, no. 12 (2023), p. 2461-2470
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- Description: We conducted a cross-sectional study to determine the prevalence of soil-transmitted helminthiases (STH) in areas of rural Alabama, USA, that have sanitation deficits. We enrolled 777 children; 704 submitted stool specimens and 227 a dried blood spot sample. We microscopically examined stool specimens from all 704 children by using Mini-FLOTAC for helminth eggs. We tested a subset by using molecular techniques: real-time PCR analysis for 5 STH species, TaqMan Array Cards for enteric helminths, and digital PCR for Necator americanus hookworm. We analyzed dried blood spots for Strongyloides stercoralis and Toxocara spp. roundworms by using serologic testing. Despite 12% of our cohort reporting living in homes that directly discharge untreated domestic wastewater, stool testing for STH was negative; however, 5% of dried blood spots were positive for Toxocara spp. roundworms. Survey data suggests substantial numbers of children in this region may be exposed to raw sewage, which is itself a major public health concern. © 2023 Centers for Disease Control and Prevention (CDC). All rights reserved.
- Authors: Poole, Claudette , Barker, Troy , Bradbury, Richard , Capone, Drew , Chatham, Amy , Handali, Sukwan , Rodriguez, Eduardo , Qvarnstrom, Yvonne , Brown, Joe
- Date: 2023
- Type: Text , Journal article
- Relation: Emerging Infectious Diseases Vol. 29, no. 12 (2023), p. 2461-2470
- Full Text:
- Reviewed:
- Description: We conducted a cross-sectional study to determine the prevalence of soil-transmitted helminthiases (STH) in areas of rural Alabama, USA, that have sanitation deficits. We enrolled 777 children; 704 submitted stool specimens and 227 a dried blood spot sample. We microscopically examined stool specimens from all 704 children by using Mini-FLOTAC for helminth eggs. We tested a subset by using molecular techniques: real-time PCR analysis for 5 STH species, TaqMan Array Cards for enteric helminths, and digital PCR for Necator americanus hookworm. We analyzed dried blood spots for Strongyloides stercoralis and Toxocara spp. roundworms by using serologic testing. Despite 12% of our cohort reporting living in homes that directly discharge untreated domestic wastewater, stool testing for STH was negative; however, 5% of dried blood spots were positive for Toxocara spp. roundworms. Survey data suggests substantial numbers of children in this region may be exposed to raw sewage, which is itself a major public health concern. © 2023 Centers for Disease Control and Prevention (CDC). All rights reserved.
Application of a universal parasite diagnostic test to biological specimens collected from animals
- Lane, Meredith, Kashani, Mitra, Barratt, Joel, Qvarnstrom, Yvonne, Yabsley, Michael, Garrett, Kayla, Bradbury, Richard
- Authors: Lane, Meredith , Kashani, Mitra , Barratt, Joel , Qvarnstrom, Yvonne , Yabsley, Michael , Garrett, Kayla , Bradbury, Richard
- Date: 2023
- Type: Text , Journal article
- Relation: International Journal for Parasitology: Parasites and Wildlife Vol. 20, no. (2023), p. 20-30
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- Description: A previously described universal parasite diagnostic (nUPDx) based on PCR amplification of the 18S rDNA and deep-amplicon sequencing, can detect human blood parasites with a sensitivity comparable to real-time PCR. To date, the efficacy of this assay has only been assessed on human blood. This study assessed the utility of nUPDx for the detection of parasitic infections in animals using blood, tissues, and other biological sample types from mammals, birds, and reptiles, known to be infected with helminth, apicomplexan, or pentastomid parasites (confirmed by microscopy or PCR), as well as negative samples. nUPDx confirmed apicomplexan and/or nematode infections in 24 of 32 parasite-positive mammals, while also identifying several undetected coinfections. nUPDx detected infections in 6 of 13 positive bird and 1 of 2 positive reptile samples. When applied to 10 whole parasite specimens (worms and arthropods), nUPDx identified all to the genus or family level, and detected one incorrect identification made by morphology. Babesia sp. infections were detected in 5 of the 13 samples that were negative by other diagnostic approaches. While nUPDx did not detect PCR/microscopy-confirmed trichomonads or amoebae in cloacal swabs/tissue from 8 birds and 2 reptiles due to primer template mismatches, 4 previously undetected apicomplexans were detected in these samples. Future efforts to improve the utility of the assay should focus on validation against a larger panel of tissue types and animal species. Overall, nUPDx shows promise for use in both veterinary diagnostics and wildlife surveillance, especially because species-specific PCRs can miss unknown or unexpected pathogens. © 2022
- Authors: Lane, Meredith , Kashani, Mitra , Barratt, Joel , Qvarnstrom, Yvonne , Yabsley, Michael , Garrett, Kayla , Bradbury, Richard
- Date: 2023
- Type: Text , Journal article
- Relation: International Journal for Parasitology: Parasites and Wildlife Vol. 20, no. (2023), p. 20-30
- Full Text:
- Reviewed:
- Description: A previously described universal parasite diagnostic (nUPDx) based on PCR amplification of the 18S rDNA and deep-amplicon sequencing, can detect human blood parasites with a sensitivity comparable to real-time PCR. To date, the efficacy of this assay has only been assessed on human blood. This study assessed the utility of nUPDx for the detection of parasitic infections in animals using blood, tissues, and other biological sample types from mammals, birds, and reptiles, known to be infected with helminth, apicomplexan, or pentastomid parasites (confirmed by microscopy or PCR), as well as negative samples. nUPDx confirmed apicomplexan and/or nematode infections in 24 of 32 parasite-positive mammals, while also identifying several undetected coinfections. nUPDx detected infections in 6 of 13 positive bird and 1 of 2 positive reptile samples. When applied to 10 whole parasite specimens (worms and arthropods), nUPDx identified all to the genus or family level, and detected one incorrect identification made by morphology. Babesia sp. infections were detected in 5 of the 13 samples that were negative by other diagnostic approaches. While nUPDx did not detect PCR/microscopy-confirmed trichomonads or amoebae in cloacal swabs/tissue from 8 birds and 2 reptiles due to primer template mismatches, 4 previously undetected apicomplexans were detected in these samples. Future efforts to improve the utility of the assay should focus on validation against a larger panel of tissue types and animal species. Overall, nUPDx shows promise for use in both veterinary diagnostics and wildlife surveillance, especially because species-specific PCRs can miss unknown or unexpected pathogens. © 2022
A pilot comparison of fixatives for hookworm real-time polymerase chain reaction
- Bradbury, Richard, Inagaki, Kengo, Singh, Gurbaksh, Agana, Urita, Patterson, Kayla, Malloch, Lacy, Rodriguez, Eduardo, Qvarnstrom, Yvonne, Hobbs, Charlotte
- Authors: Bradbury, Richard , Inagaki, Kengo , Singh, Gurbaksh , Agana, Urita , Patterson, Kayla , Malloch, Lacy , Rodriguez, Eduardo , Qvarnstrom, Yvonne , Hobbs, Charlotte
- Date: 2023
- Type: Text , Journal article
- Relation: American Journal of Tropical Medicine and Hygiene Vol. 108, no. 2 (2023), p. 335-339
- Full Text:
- Reviewed:
- Description: Polymerase chain reaction (PCR) is increasingly used in the diagnosis of soil-transmitted helminth infections. Despite this, few studies have evaluated the impact of different fecal fixatives on the outcome of fecal helminth qPCR analysis, and none have evaluated the effect of commercial parasitology fixatives commonly used in diagnostic laboratories. We fixed dog feces containing Ancylostoma spp. hookworm eggs in zinc polyvinyl alcohol (Zn-PVA) and Total-Fix, and with 70% ethanol (EtOH) as a control. DNA was extracted at timepoints 11, 33, 64, and 94 days and subjected to Ancylostoma spp. quantitative PCR (qPCR). A linear regression model was created to assess the effect of preservative types on the temporal change of qPCR quantification cycle number (Cq) values, accounting for variances among individual animals. Fixation in 70% EtOH least affected Cq values over 94 days. Total-Fix preservation yielded a higher Cq overall, but there was no significant difference compared with 70% EtOH fixation. Fixation in Zn-PVA resulted in significantly (P < 0.001) higher Cq values than 70% EtOH after only 33 days and loss of amplification at 64 days. Consistent with other helminth fixation studies, 70% EtOH performed well in preserving hookworm DNA over 94 days. Total-Fix provided a comparable alternative for qPCR analysis for hookworm. Fixation in Zn-PVA resulted in loss of detectable hookworm DNA at 64 days, as determined by qPCR. Copyright © 2023 The American Society of Tropical Medicine and Hygiene.
- Authors: Bradbury, Richard , Inagaki, Kengo , Singh, Gurbaksh , Agana, Urita , Patterson, Kayla , Malloch, Lacy , Rodriguez, Eduardo , Qvarnstrom, Yvonne , Hobbs, Charlotte
- Date: 2023
- Type: Text , Journal article
- Relation: American Journal of Tropical Medicine and Hygiene Vol. 108, no. 2 (2023), p. 335-339
- Full Text:
- Reviewed:
- Description: Polymerase chain reaction (PCR) is increasingly used in the diagnosis of soil-transmitted helminth infections. Despite this, few studies have evaluated the impact of different fecal fixatives on the outcome of fecal helminth qPCR analysis, and none have evaluated the effect of commercial parasitology fixatives commonly used in diagnostic laboratories. We fixed dog feces containing Ancylostoma spp. hookworm eggs in zinc polyvinyl alcohol (Zn-PVA) and Total-Fix, and with 70% ethanol (EtOH) as a control. DNA was extracted at timepoints 11, 33, 64, and 94 days and subjected to Ancylostoma spp. quantitative PCR (qPCR). A linear regression model was created to assess the effect of preservative types on the temporal change of qPCR quantification cycle number (Cq) values, accounting for variances among individual animals. Fixation in 70% EtOH least affected Cq values over 94 days. Total-Fix preservation yielded a higher Cq overall, but there was no significant difference compared with 70% EtOH fixation. Fixation in Zn-PVA resulted in significantly (P < 0.001) higher Cq values than 70% EtOH after only 33 days and loss of amplification at 64 days. Consistent with other helminth fixation studies, 70% EtOH performed well in preserving hookworm DNA over 94 days. Total-Fix provided a comparable alternative for qPCR analysis for hookworm. Fixation in Zn-PVA resulted in loss of detectable hookworm DNA at 64 days, as determined by qPCR. Copyright © 2023 The American Society of Tropical Medicine and Hygiene.
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