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MSphere 2018Human papillomavirus (HPV) vaccination elicits high-titer genotype-specific antibody responses that are associated with a reduced risk of cervical disease caused by... (Comparative Study)
Comparative Study
Human papillomavirus (HPV) vaccination elicits high-titer genotype-specific antibody responses that are associated with a reduced risk of cervical disease caused by vaccine-incorporated genotypes. Our objective was to evaluate dried blood spots (DBSs) and oral mucosal transudate (OMT) as alternative samples to serum to confirm HPV vaccine antibody status. A study was carried out to evaluate the feasibility of detecting HPV16 and HPV18 antibodies in OMT, DBSs, and sera among women who self-reported being unvaccinated or fully vaccinated with the HPV vaccine. Serum had the highest sensitivity (100%) for detection of antibodies against both HPV16 and HPV18 but the lowest specificity, due to the detection of natural infection antibodies in 16% of unvaccinated women. Conversely, DBSs and OMT had lower sensitivity (96% and 82%, respectively) but high specificity (98%). We confirmed that these antibodies were functional (i.e., neutralizing) and that their detection was quantitatively reproducible and well correlated between sample types when normalized to IgG content. DBSs and OMT are appropriate alternative sample types for HPV vaccine surveillance. These alternative sample types warrant consideration for the purposes of cervical screening, diagnosis, and management, but more work will be needed to establish the stringent parameters required for such application. Human papillomavirus (HPV) is the causative agent of cervical and other anogenital cancers. HPV vaccination, primarily targeted at young girls before the age of sexual debut, is starting to demonstrate population-level declines in HPV infection and early disease associated with vaccine-incorporated genotypes. Monitoring young women for vaccine-specific antibody is important for vaccine surveillance and may be useful as an adjunct test within a cervical screening context. We evaluated serum, dried blood spots, and oral fluid as potential samples for such applications and report robust measures of diagnostic accuracy. This is the first time a direct comparison of alternative sample types has been made between vaccinated and unvaccinated women for the detection and quantitation of HPV antibodies.
Topics: Antibodies, Viral; Desiccation; Epidemiological Monitoring; Humans; Mouth Mucosa; Papillomavirus Infections; Sensitivity and Specificity; Specimen Handling
PubMed: 29743199
DOI: 10.1128/mSphere.00043-18 -
HIV Medicine Aug 2008Dried sample spots have molecular applications, but few data are available on conditions of HIV-1 RNA amplification.
BACKGROUND
Dried sample spots have molecular applications, but few data are available on conditions of HIV-1 RNA amplification.
OBJECTIVES
To determine the impact of (i) the sample type (plasma or serum), (ii) various storage periods, (iii) transfer at ambient temperature of the dried spots via postal mail, and (iv) two different methods of elution-extraction, to amplify partial pol and env genes with a view to both phylogenic and resistance analyses.
METHODS
Fourteen samples (dried plasma spot and dried serum spot) from seven patients were stored at 20-25 degrees C for 2, 5 and 7 days. Two extraction buffers were tested on these samples, followed by reverse transcriptase-polymerase chain reaction (RT-PCR) on pol and env genes. Sixteen spots from eight other patients were sent by postal mail at ambient temperature between two laboratories, and were analysed at both sites.
RESULTS
We observed no influence of the 2-7 day storage period, whatever the sample type and viral load, but the choice of the extraction procedure is a critical step. Analysis of the mailed spots resulted in a good agreement between the two laboratories.
CONCLUSIONS
This study shows that amplification of HIV-1 RNA on spots is possible in various conditions by different operators and using appropriate reagents. This finding could be particularly useful for large-scale molecular and resistance epidemiological studies in resource-rich and resource-limited countries alike.
Topics: Blood; Blood Preservation; Desiccation; Drug Resistance, Viral; HIV Infections; HIV-1; Humans; RNA, Viral; Reverse Transcriptase Polymerase Chain Reaction; Specimen Handling; Temperature; Time Factors
PubMed: 18557950
DOI: 10.1111/j.1468-1293.2008.00604.x -
Journal of Diabetes Science and... Sep 2009Glycated hemoglobin A1c (A1C) is an important marker in the diagnosis and treatment of diabetes. Dried blood measurement of A1C is useful in large scale epidemiological... (Comparative Study)
Comparative Study
BACKGROUND
Glycated hemoglobin A1c (A1C) is an important marker in the diagnosis and treatment of diabetes. Dried blood measurement of A1C is useful in large scale epidemiological evaluation of A1C, especially to assess the impact of intervention programs. The possibility of using dried blood for measurement of A1C by the immunoturbidimetric method was explored in the present study.
METHOD
Blood was collected from 30 patients, and blood spots were prepared and dried. The dried blood spot samples were kept for different lengths of time at 4 degrees C to assess stability. Glycated hemoglobin was measured in whole blood and dried blood on the day of collection as well as on days 10 and 15 by immunoturbidimetric method.
RESULTS
The A1C values of 30 samples analyzed for comparison between whole blood estimation and dried blood ranged from 4.6% to 9.9%. The mean A1C on the day of sample collection was 6.01% +/- 1.58% in fresh whole blood samples and 5.94% +/- 1.58 % in dried blood spots. A linear and highly correlated relationship was observed between dried blood A1C values and those in whole blood (r = 0.986 and intraclass correlation value = 0.993). Glycated hemoglobin values on day 10 and day 15 were comparable with the values on day 1 with a shift in mean of just 1% on day 10 and 3.04% on day 15.
CONCLUSION
In conclusion, dried blood can be used for measurement of A1C by immunoturbidimetric method, and further stability of A1C measurement from dried blood for up to 15 days at 4 degrees C makes it an ideal matrix for transportation in developing countries like India.
Topics: Biomarkers; Blood Specimen Collection; Glycated Hemoglobin; Humans; Immunoassay; Nephelometry and Turbidimetry; Protein Stability; Reproducibility of Results; Temperature; Time Factors
PubMed: 20144437
DOI: 10.1177/193229680900300527 -
Journal of Clinical Microbiology Dec 2020The treatment of HIV-2 in resource-limited settings (RLS) is complicated by the limited availability of HIV-2-active antiretroviral drugs and inadequate access to HIV-2...
The treatment of HIV-2 in resource-limited settings (RLS) is complicated by the limited availability of HIV-2-active antiretroviral drugs and inadequate access to HIV-2 viral load and drug resistance testing. Dried blood spots (DBS)-based drug resistance testing, widely studied for HIV-1, has not been reported for HIV-2 and could present an opportunity to improve care for HIV-2-infected individuals. We selected 150 DBS specimens from ongoing studies of antiretroviral therapy (ART) for HIV-2 infection in Senegal and subjected them to genotypic drug resistance testing. Total nucleic acid was extracted from DBS, reverse transcribed, PCR amplified, and analyzed by population-based Sanger sequencing, and major drug resistance-associated mutations (RAM) were identified. Parallel samples from plasma and peripheral blood mononuclear cells (PBMC) were also genotyped. We obtained 58 protease/reverse transcriptase genotypes. Plasma viral load was significantly correlated with genotyping success ( < 0.001); DBS samples with corresponding plasma viral load >250 copies/ml had a success rate of 86.8%. In paired DBS-plasma genotypes, 83.8% of RAM found in plasma were also found in DBS, and replicate DBS genotyping revealed that a single test detected 86.7% of known RAM. These findings demonstrate that DBS-based genotypic drug resistance testing for HIV-2 is feasible and can be deployed in RLS with limited infrastructure.
Topics: Drug Resistance, Viral; Genotype; HIV Infections; HIV-2; Humans; Leukocytes, Mononuclear; Senegal; Specimen Handling; Viral Load
PubMed: 33055182
DOI: 10.1128/JCM.02303-20 -
Emerging Infectious Diseases Dec 2023We 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 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.
Topics: Child; Animals; Humans; Cross-Sectional Studies; Soil; Alabama; Helminthiasis; Helminths; Feces; Prevalence
PubMed: 37987581
DOI: 10.3201/eid2912.230751 -
American Journal of Human Biology : the... Jan 2018Telomere length (TL) is a biomarker of aging and age-related decline. Although venous blood is considered the "gold standard" for TL measurement, its collection is often... (Comparative Study)
Comparative Study
OBJECTIVES
Telomere length (TL) is a biomarker of aging and age-related decline. Although venous blood is considered the "gold standard" for TL measurement, its collection is often not feasible or desired in nonclinical settings. Saliva and dried blood spots (DBS) have been used as alternatives when venipuncture cannot be performed. However, it is not known whether these sample types yield TL measurements comparable to those obtained from venous blood. We sought to determine whether different samples from the same individual yield comparable TL measurements.
METHODS
We extracted DNA from matched buffy coat, saliva (Oragene and Oasis), and DBS (venous and capillary) samples from 40 women aged 18-77 years. We used the monochrome multiplex qPCR (MMQPCR) assay to measure TL in all sample types for each participant and applied quality control measures to retain only high-quality samples for analysis. We then compared TL from buffy coat and saliva to examine how these measurements differ and to test if TL is correlated across sample types.
RESULTS
TL differed significantly across buffy coat, Oragene saliva, and Oasis saliva samples. TL from buffy coat and Oragene saliva was moderately correlated (ρ = 0.48, P = .002) and the most similar in size. Oasis saliva TL was not correlated with buffy coat or Oragene saliva TL, and was the shortest. DBS DNA yields were inadequate for TL measurement using the MMQPCR assay.
CONCLUSIONS
Using a matched dataset we demonstrate that sample type significantly influences the TL measurement obtained using the MMQPCR assay.
Topics: Blood Chemical Analysis; Dried Blood Spot Testing; Multiplex Polymerase Chain Reaction; Saliva; Specimen Handling; Telomere
PubMed: 28949426
DOI: 10.1002/ajhb.23062 -
F1000Research 2023The sternum exhibits unique anatomical variations with major clinical and forensic implications. This study is devoted to providing baseline epidemiological information... (Observational Study)
Observational Study
The sternum exhibits unique anatomical variations with major clinical and forensic implications. This study is devoted to providing baseline epidemiological information about the sternal foramen and variant xiphoid morphology in Ethiopia. Two extremely interesting and unusual variations of the sternal foramen are also discussed. This observational study was carried out using dried adult human sternum obtained from skeletal remains samples brought for medicolegal examination over a period of 4 years. A total of 94 dried adult human sternums (66 males (70.2%) and 28 females (29.8%)) were obtained with an age range of 21 to 57 years and a mean age at death of 38.383 ± 11.3480 years. Dried human sternum specimens were morphologically examined, and morphometric parameters were recorded and photographed. A sternal foramen was found in 18 specimens (19.1%); 17 were male and one was female. A single sternal foramen was observed in 83.3% (n=15/18) of the sternal bodies and 11.1% (n=2/18) of the xiphoid processes (both males). In addition, a double sternal foramen was observed in a single male specimen on the mesosternum and xiphoid process. The most common sternal foramen site was at the fifth costochondral junction level. The xiphoid process was present in 77 samples and ended as a single process in 83.1% (n=64/77) of samples. In 15.6% (n=12/77) of the samples, the xiphoid process was bifurcated and trifurcated in a single male (1.3%) specimen. The sternal foramen and variation in xiphoid morphology are common anatomical variations in Ethiopia. The findings of the current study highlight the necessity of strict precautionary measures during sternal procedures in this study population. In addition, such incidental findings during radiologic and autopsy procedures should be properly evaluated to avoid misdiagnosis and misinterpretation of such findings as traumatic or pathologic conditions.
Topics: Humans; Sternum; Male; Female; Ethiopia; Adult; Middle Aged; Young Adult; Anatomic Variation; Xiphoid Bone
PubMed: 38845617
DOI: 10.12688/f1000research.133151.1 -
Journal of Mass Spectrometry and... Apr 2021Inborn errors of propionate, cobalamin and methionine metabolism are targets for Newborn Screening (NBS) in most programs world-wide, and are primarily screened by...
Analysis of 2-methylcitric acid, methylmalonic acid, and total homocysteine in dried blood spots by LC-MS/MS for application in the newborn screening laboratory: A dual derivatization approach.
Inborn errors of propionate, cobalamin and methionine metabolism are targets for Newborn Screening (NBS) in most programs world-wide, and are primarily screened by analyzing for propionyl carnitine (C3) and methionine in dried blood spot (DBS) cards using tandem mass spectrometry (MS/MS). Single-tier NBS approaches using C3 and methionine alone lack specificity, which can lead to an increased false-positive rate if conservative cut-offs are applied to minimize the risk of missing cases. Implementation of liquid chromatography tandem mass spectrometry (LC-MS/MS) second-tier testing for 2-methylcitric acid (MCA), methylmalonic acid (MMA), and homocysteine (HCY) from the same DBS card can improve disease screening performance by reducing the false-positive rate and eliminating the need for repeat specimen collection. However, DBS analysis of MCA, MMA, and HCY by LC-MS/MS is challenging due to limited specimen size and analyte characteristics leading to a combination of low MS/MS sensitivity and poor reverse-phase chromatographic retention. Sufficient MS response and analytical performance can be achieved for MCA by amidation using DAABD-AE and by butylation for MMA and HCY. Herein we describe the validation of a second-tier dual derivatization LC-MS/MS approach to detect elevated MCA, MMA, and HCY in DBS cards for NBS. Clinical utility was demonstrated by retrospective analysis of specimens, an interlaboratory method comparison, and assessment of external proficiency samples. Imprecision was <10.8% CV, with analyte recoveries between 90.2 and 109.4%. Workflows and analytical performance characteristics of this second-tier LC-MS/MS approach are amenable to implementation in the NBS laboratory.
PubMed: 34820666
DOI: 10.1016/j.jmsacl.2021.03.001 -
Frontiers in Immunology 2023Among the challenges in schistosomiasis surveillance and mapping surveys is the lack of a sensitive diagnostic method especially in low transmission setting. Currently,...
BACKGROUND
Among the challenges in schistosomiasis surveillance and mapping surveys is the lack of a sensitive diagnostic method especially in low transmission setting. Currently, the WHO recommends the use point-of-care circulating cathodic antigen (Schisto POC-CCA) tests for surveillance and mapping of intestinal schistosomiasis. However, Schisto POC-CCA test has its drawbacks, one of which is the timely availability of test kits. One approach to overcoming this challenge is to develop a low-cost sampling method that allows for the collection and transport of urine specimens even in resource-limited settings.
OBJECTIVE
To develop a simple and efficient method for the collection and detection of () CCA using urine spotted onto filter paper.
METHODOLOGY
To develop a dried urine spot (DUS) method, various dried matrix extraction parameters were tested and optimized using predesigned steps. The parameters include the size of filter paper (determined by the number of punches), volume of solvents, and type of solvent. Moreover, we optimized the incubation conditions (time and temperature). Urine and stool specimens to conduct the experiments were collected from volunteer fishermen in Mwanza and this project staff. Data were entered into the Microsoft Excel spreadsheet and IBM Statistical Package for the Social Sciences, version 20 for analysis.
RESULTS
The optimal results were obtained when the procedure was run under the following conditions: Five punches of filter paper containing DUS were dissolved in 150 µl of distilled water and incubated at room temperature for 24 hours in an Eppendorf tube. More than 93% of the assays performed under these conditions produced results that were either comparable to or significantly better than the standard method.
CONCLUSION
This study demonstrates the feasibility of collecting urine specimen (DUS) using filter paper and detecting CCA from DUS specimen using the Schisto POC-CCA cassette test.
Topics: Humans; Animals; Schistosoma mansoni; Proof of Concept Study; Sensitivity and Specificity; Antigens, Helminth; Tanzania; Antibodies, Helminth
PubMed: 37753086
DOI: 10.3389/fimmu.2023.1216710 -
Cancer Cytopathology May 2017The Royal College of Pathologists of Australasia Cytopathology Quality Assurance Program has operated an external quality assurance program in nongynecologic...
BACKGROUND
The Royal College of Pathologists of Australasia Cytopathology Quality Assurance Program has operated an external quality assurance program in nongynecologic cytopathology since 1993. Glass slide preparations of a wide range of nongynecologic cases were circulated to approximately 200 cytopathology laboratories in 16 countries.
METHODS
General nongynecologic cytology cases were manufactured from residual specimens after routine diagnosis. Fine-needle aspiration (FNA) cases were made by sampling fresh tissue and making direct specimens. The majority of cases consisted of both air-dried and fixed preparations. Results returned to laboratories included illustrated case discussions highlighting diagnostic features, key differential diagnoses, and useful adjunctive tests.
RESULTS
The current study reviewed >22,000 results for 123 nongynecologic cases. Cases found to cause the most diagnostic difficulties included serous effusion cases with metastatic carcinoma in a dispersed pattern, well-differentiated carcinoma, and cellular reactive cases; urine specimens with sparse malignant cells; reactive pneumocytes in a bronchoalveolar lavage; breast FNA cases with papillary lesions; gestational specimens; and fibroadenoma. FNA specimens from the lung and thyroid, particularly papillary thyroid carcinoma, generally were well reported.
CONCLUSIONS
The use of multiple preparations of the same specimen has allowed interlaboratory comparison, and the quality assurance program has played an educational role as well as informing the laboratory accreditation process. Cancer Cytopathol 2017;125:349-361. © 2017 American Cancer Society.
Topics: Adenocarcinoma; Ascitic Fluid; Australasia; Biopsy, Fine-Needle; Body Fluids; Breast Neoplasms; Bronchoalveolar Lavage Fluid; Carcinoid Tumor; Carcinoma; Carcinoma, Non-Small-Cell Lung; Carcinoma, Papillary; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cytodiagnosis; Fibroadenoma; Humans; Leukemia, Myeloid, Acute; Lung Neoplasms; Melanoma; Mesothelioma; Neoplasms; Neoplasms, Cystic, Mucinous, and Serous; Pathology, Clinical; Pericardial Fluid; Quality Assurance, Health Care; Small Cell Lung Carcinoma; Thyroid Cancer, Papillary; Thyroid Neoplasms; Urine
PubMed: 28241108
DOI: 10.1002/cncy.21838