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:
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- 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.
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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- 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.
- 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.
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