Original Research

Efficacy of integrated rapid diagnostic testing for HIV, syphilis and hepatitis in Zimbabwe

Newten Handireketi, Pisirai Ndarukwa, Tonderai Manengureni, Loice Cushny, Moses J. Chimbari

Received: 01 Apr. 2025; Accepted: 25 Sept. 2025; Published: 09 Dec. 2025

Copyright: © 2025. The Authors. Licensee: AOSIS.
This work is licensed under the Creative Commons Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/).

Abstract

Background: Triple point-of-care tests for simultaneous detection of human immunodeficiency virus (HIV), syphilis and hepatitis B virus (HBV) make service delivery simple, reduce costs and time for testing, and lower supply chain barriers. The performance of such antenatal care (ANC) panel has not been evaluated in Zimbabwe.

Aim: The aim of the study was to compare the performance of the ANC panel with standard testing.

Setting: The study was carried out in Mudzi District, Zimbabwe.

Methods: An analytical cross-sectional design was followed to compare the performance of HIV/syphilis/HBV surface antigen (HBsAg) rapid diagnostic tests against standard of screening done using individual testing devices. Reference tests had HIV and syphilis on one strip and HBV on a separate strip. Data were collected between May and October 2024 at nine clinics and one hospital. In all, 284 participants took part in the study. Informed consent was obtained before data collection. A finger-prick sample was tested. Sensitivity, specificity, negative predictive value (NPV) and positive predictive value (PPV) were determined against routine testing.

Results: The results were 100.00% for HIV and syphilis on sensitivity, specificity, PPV and NPV. The HIV and syphilis proportions were 2.46% and 2.11%, respectively. Specificity and NPV for HBV were 100.00%. Sensitivity and PPV for HBV were not evaluated because there were no positive cases. Overall, the panel and routine testing had same results across all the three infections.

Conclusion: The panel produced the same efficacy as routine care for simultaneous screening of the infections and is recommended for point-of-care use.

Contribution: This evidence supports effectiveness of integrated testing as an alternative to routine care.

Keywords: antenatal care panel; HIV; syphilis; hepatitis B; integration; triple elimination; efficacy; public health.

Introduction

Human immunodeficiency virus (HIV), syphilis and viral hepatitis are major global public health problems that can be transmitted from mother-to-child. The three unilaterally and jointly contribute to significant morbidity and mortality, and are therefore targeted for elimination through an integrated approach.¹ In 2023, women and girls accounted for 44.0% of 1.3 million (1 million – 1.7 million) HIV incidence worldwide.² In the same year, the HIV global prevalence among pregnant women was about 1.2 million (950 000 – 1.4 million) according to World Health Organization (WHO) estimates,³ while an estimated 50 000 new infections from vertical transmission were reported by the United Nations International Children’s Emergency Fund (UNICEF).⁴ Globally, syphilis accounts for about 2 million cases in pregnant women, around 460 000 new cases of miscarriages and stillbirth, 270 000 incidences of congenital syphilis and 270 000 cases of low birth weight.⁵ About 65 million women of reproductive age groups have hepatitis B virus (HBV), with 9 out of every 10 infected mothers transmitting the virus to their children perinatally.⁶ High burden and co-morbidity from these infections have been recorded in sub-Saharan Africa, including Zimbabwe.^7,8,9 Previous results from Zimbabwe showed that about 45.5% of females had active syphilis co-occurring with HIV. This likelihood is 3.7 (adjusted odds ratio [aOR]: 3.7, 95% confidence interval [CI]: 2.3–5.9) times higher in females living with HIV than in HIV-negative females.¹⁰ In the same Zimbabwean setting, hepatitis B surface antigen (HBsAg)-seroprevalence of 4.11% (95% CI: 2.52, 5.68) has been reported among HBsAg/HIV-coinfected pregnant women who were at least 20 weeks’ gestational age.¹¹ Zimbabwe adopted the WHO targets for eliminating HIV, syphilis and viral hepatitis as public health threats by 2030.¹²

Laboratory-based diagnostic tests are often unavailable for populations living in areas where they are needed most, particularly in primary healthcare facilities in low- and middle-income countries.¹³ This highlights the importance of accessibility to simple and affordable tests with shorter time to results such as rapid diagnostic tests (RDTs). The introduction of such point-of-care (POC) testing in Zimbabwe has made patient care and the prevention of mother-to-child transmission (PMTCT) services widely available.¹⁴ Rapid diagnostic test devices are widely used to screen for different infections such as HIV, syphilis and HBV. The modes of transmission for HIV, syphilis and HBV overlap, providing opportunities for integrated control interventions such as the use of triple point-of-care tests (POCTs). Infection with HIV can accelerate the course of concurrent syphilis and viral hepatitis (B & C) disease progression. Possible strategies to achieve triple elimination of the three infections in sub-Saharan Africa include scaling up the integrated screening within antenatal care (ANC) settings to help improve coverage and guide treatment.¹⁵ Integrated testing services refer to simultaneous testing of several infectious diseases in primary health care settings and this approach is feasible, with the ability to detect infections before they are deadly.¹⁶ Implementing integrated HIV, viral hepatitis and sexually transmitted diseases (STDs) services has also been demonstrated to increase access, improve timeliness of the service delivery and increase effectiveness of efforts to prevent the infections that share common host risk factors, behaviours and social determinants.¹⁷ However, available diagnostics frequently vary in quality.¹⁸ The performance of an RDT is critical in providing accurate screening for infections in places near or at the POC and further patient management. The performance of RDTs is determined by calculating sensitivity, specificity and predictive values when compared with a reference method, which is considered a ‘gold standard’. The sensitivity and specificity values reported by manufacturers may be unreliable, so independent studies and/or strict regulatory approvals are critical to determine the true values in real-world settings before use.¹⁹

Available evidence shows that the performance characteristics of RDTs for integrated testing of HIV, syphilis and HBV have not been evaluated in Zimbabwe. Evaluation of such a panel’s efficacy is very relevant, as there is no solid evidence for its use on the Zimbabwean population. Hence, our study aimed to provide such evidence by assessing the triple POCT’s efficacy.

Research methods and design

Study design

An analytical cross-sectional study in which data were collected from study participants reporting to the healthcare facilities for ANC services was conducted between May 2024 and October 2024.

Study setting

This study was done in Mudzi district, Mashonaland East Province, Zimbabwe, at nine clinics and one hospital that provided primary healthcare services.

Study population

This study involved pregnant women who presented at healthcare facilities for their routine ANC and other clinical services. Participants aged 18 years or older and emancipated minors who sought ANC services were recruited for the study.

Sample size and sampling procedure

Sample size was determined using Australian Emergency Veterinary (AUSVET) for a true prevalence with an imperfect test, assuming:

Human immunodeficiency virus

• True prevalence = 0.12²⁰
• Sensitivity = 0.95
• Specificity = 0.95
• Desired precision = 0.05
• Confidence = 0.95

Human immunodeficiency virus: The minimum sample size was 253 at the analysis stage. Assuming 12% attrition, the desired sample size therefore was 284 pregnant women.

Syphilis

• True prevalence = 0.02²¹
• Sensitivity = 0.95
• Specificity = 0.95
• Desired precision = 0.05
• Confidence = 0.95

Syphilis: The minimum sample size was 121 at the analysis stage. Assuming 12% attrition, the desired sample size therefore was 136 pregnant women.

Hepatitis B virus

• True prevalence = 0.08²²
• Sensitivity = 0.95
• Specificity = 0.95
• Desired precision = 0.05
• Confidence = 0.95

Hepatitis: The minimum sample size was 204 at the analysis stage. Assuming 12% attrition, the desired sample size therefore is 229 pregnant women.

Therefore, the sample size calculation was based on estimating the prevalence of HIV (12%), which gives the highest sample size (284) relative to the calculations for syphilis and HBV.

A total of 284 participants (N = 284) were recruited for the study. Systematic random sampling was employed in selecting participants for the study. The first individual who came to the healthcare facility was not selected to participate in the study; the second one was selected, and subsequently, the participants with odd numbers were not included. The healthcare facilities were identified by selecting the first 10 high-volume sites for antenatal clinic services in the district.

Study procedure

Each patient initially underwent testing for HIV, syphilis, and HBV following routine standard operating procedures and algorithms, after which the integrated ANC panel test was also performed on the same individual. Informed consent was obtained from all participants who took part in the study prior to data collection. According to the standard of care National Rapid testing algorithm in Zimbabwe (Appendix 1)²³, all pregnant women were tested for HIV and syphilis on one RDT testing strip, a duo testing device A1, and then screened for HBV separately (Appendix 2).²⁴

Antenatal care Panel testing was performed according to the instructions provided by Abbott as detailed in the Standard Operating Procedure (Appendix 3).²⁵ All healthcare workers involved in the project were familiarised with the Standard Operating Procedure during a 5-day workshop conducted by the Medical Laboratory and Clinical Scientists Council of Zimbabwe. For this purpose, 0.2 mL of finger-prick capillary whole blood specimens were collected from each participant using ethylenediaminetetraacetic acid (EDTA) capillary blood collection tubes. The finger-prick capillary whole blood specimens were immediately tested on the ANC Panel test, and the results were recorded.

Precisely, 50 µL of blood sample collected from the small EDTA capillary blood collection tubes with 0.2 ml of the specimens were applied to the sample pad (within the sample application area) of each strip on the panel. A drop of chase buffer was applied 1 min after the sample. The results were interpreted at 25 min after sample application.

Capillary (finger-prick) whole blood specimens were easy to collect and use in the field, as they did not require a separate storage container or expert for blood collection. The results from routine care were compared with results from ANC panel.

Data were entered on Open Data Kit (ODK), a collection of open-source and free software for collecting, managing and using data under resource-constrained environments. Consolidated data were downloaded as an Excel file. The Excel file was then imported into STATA version 14.0 for analysis.

Ethical considerations

Ethical approval to conduct this study was obtained from the Medical Research Council of Zimbabwe (MRCZ) (No. MRCZ/A/3106) and the University of KwaZulu-Natal Biomedical Research Ethics Committee (No. BREC/00006499/2023). Written informed consent was obtained from the participants of the study. All procedures performed in this study were in accordance with ethical standards of the MRCZ, BREC, the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

Results

Demographic characteristics

Data analysis was done for 284 participants. The median and interquartile range (IQR) for age of the participants was 25 years (IQR: 20–31 years). The majority of them (67.6%, n = 192/284) had a grade seven to ordinary level education qualification. The demographic characteristics of participants who were tested using the Abbott ANC panel are presented in Table 1.

Proportions of participants with human immunodeficiency virus, syphilis and hepatitis B infection

The proportions for HIV and syphilis infection in the sample were 2.5% (n = 7/284; 95% CI: 1.0–5.0) and 2.1% (n = 6/284; 95% CI: 0.78–4.54), respectively. The proportion of HIV and syphilis co-infection was 0.7% (n = 2/284). The proportion of any sexually transmitted infection (STI) (HIV or syphilis) was 3.9% (n = 11/284; 95% CI: 1.95–6.82). None of the participants had HBV infection.

Sensitivity, specificity and predictive values of the antenatal care panel in diagnosing human immunodeficiency virus, syphilis and hepatitis B infection

For HIV, the sensitivity and specificity were 100.0% (n = 7/7) and 100.0% (n = 277/277), respectively. The positive and negative predictive values (NPVs) were also 100.0% (n = 7/7) and 100.00% (n = 277/277), respectively. For syphilis, the sensitivity and specificity were 100.0% (n = 6/6) and 100.0% (n = 278/278), respectively, and the positive and NPVs were 100.0% (n = 6/6) and 100.0% (n = 278/278), respectively. For HBV, the specificity and NPV were 100.0% (n = 284/284) and 100.0% (n = 284/284), respectively. Sensitivity and positive predictive values (PPVs) could not be calculated as there were no cases detected. The results for the performance of the ANC panel compared to routine care, the gold standard, are shown in Table 2.

Discussion

The WHO recommends testing pregnant women for HIV, syphilis and HBV to achieve triple elimination of their mother-to-child transmissions,²⁶ but progress has been slow in sub-Saharan Africa, which bears the highest burden of the three diseases. Point-of-care tests can be used for testing of pregnant women as part of ANC.²⁷ Guidelines for POCTs emphasise the need for highly sensitive and specific tests to support informed clinical decision-making.²⁸ This study compared the performance characteristics of an ANC panel kit for integrated testing of HIV, syphilis and HBV with results from standard of care under a non-laboratory setting. Recommendations have been made for conducting real-life evaluations of diagnostic tests in population where the tests will be used. Such evaluations provide clinically accurate data appropriate for the local prevalence and for other context-specific factors that could influence their accuracy, such as common comorbidities.²⁹ We found that the performance characteristics of the ANC panel for integrated testing of HIV, syphilis and HBV were the same as the standard of care. Although no samples tested positive for HBV by both integrated testing and the standard of care, our results demonstrated that the integrated approach brought hepatitis testing to the same level as HIV and syphilis. In the current standard of care, HIV and syphilis are tested simultaneously with blood sample collected from one prick. Hepatitis B virus is either tested separately, or not tested at all with patients referred to the district hospital. The algorithm does not make it mandatory to test for HBV at the same time as HIV and syphilis. The integrated approach allowed every pregnant woman who was tested for HIV had syphilis to be tested for HBV as well. The tests for all the three infections using a blood sample from one finger-prick were enough for the three testing devices, which were bundled in one kit for integrated triple testing.

Results from this study agree with previous studies from a systematic review and meta-analysis of 18 studies that evaluated three dual HIV and syphilis POCTs. The review showed sensitivity and specificity of 93.8% – 100% and 94.2% – 100% respectively for HIV, with sensitivity and specificity of 47.4% – 100% and 90.8% – 100% respectively for syphilis.³⁰ Our results concur with findings on evaluations done with individual test strips of the panel in reports released by WHO. The reports showed (1) sensitivity of 100% for both HIV and syphilis testing, and (2) specificity of 99.4%, 100% and 100% for HIV, syphilis and HBV, respectively,³¹ comparable to 100% from this study in each case. The results of HIV and syphilis co-infection in a proportion of 0.7% (n = 2) of the entire sample and 28.6% among all (n = 7) of those with HIV also relate to previous findings in Zimbabwe which showed people living with HIV being more likely to have active syphilis (aOR: 2.2; 95% CI: 1.4–3.6).³² Co-infection between HIV and syphilis supports the need for integrated approach to individuals’ STIs testing and treatments, as recommended by previous findings and the WHO, in health delivery points such as ANC settings.¹⁰ The high sensitivity and specificity minimise false negatives and false positives respectively.

The low number of HIV-positive patients (n = 7; 2.5%) for both the integrated approach and standard of care may be explained by the decline in new HIV infections for all age groups as indicated by incidence of about 0.17% in 2022 for the country.³³ The PPV and sensitivity calculations were based on the 7 positive results. The specificity of the integrated approach for HIV testing, which was 100%, showed that there were no false positives in this sample. Also, the sensitivity of the integrated approach for HIV, which was 100%, showed that there were no false negatives.

The syphilis proportion of 2.11% (95% CI: 0.78–4.54) among the 284 patients we tested with both the integrated approach and standard of care compares favourably with a prevalence of 1% obtained in another study that was carried out in Zimbabwe.¹⁰ The PPV and sensitivity calculations were based on the 6 positive results. The PPV, sensitivity and specificity of the ANC panel were high (100%) in each case for syphilis. This showed perfect accuracy compared with the standard of care, with no false positives and false negatives. This makes the integrated testing useful for syphilis diagnostic purposes in ANC settings.

All of the 284 patients tested for HBV were negative with both ANC panel and standard of care. Therefore, sensitivity and PPV could not be evaluated. The discrepancy between high burden and co-morbidity with HIV and other STI reported in previous studies may be explained by several factors. This observed difference may be attributed to geographical variability in HBV prevalence. This is the first study that has been carried out in this setting, determining the proportion of pregnant women infected by HBV. Results from 21 previous studies comprising 291 784 persons tested for HBsAg revealed a low age-adjusted prevalence rate of 0.65% (95% CI: 0.31% – 1.00%) in blood donors showing decreasing prevalence of HBV.³⁴ However, lower results from this study may be explained by differences in study populations that were targeted. Women attending ANC services receive preventive interventions against STIs in general, and that reduces chances of getting HBV. The results may be because of chance, and population-based surveys may produce better estimates of the true burden. The specificity of the Abbott ANC panel diagnostic test for viral hepatitis, which was 100%, showed that there were no false positives. The ANC panel demonstrated that it performed the same way with the standard of care on HBV testing among those without the infection. In spite of the absence of reactive HBV results from this sample, precautions during antenatal period remain critical considering the risk of vertical transmission.

The limitations of results from this study were because of the low prevalence of HIV, syphilis and HBV. Although all efforts were taken to collect blood for testing from a random sample, random sampling error could have introduced variations into the results by chance. The study was carried out only among those who came to receive ANC services at primary healthcare facilities and may not represent those who do not access ANC services.

Overall, the results of the study demonstrated that integrated testing for HIV, syphilis and HBV has the same efficacy with standard of care. However, we place a caveat on the results that they need to be interpreted, noting that the number of positives was low for HIV as well as syphilis, and there were no positives for HBV in this sample. The public health implications of adopting this approach have demonstrated that, in most studies among all assessed settings, comprehensive, population-based antenatal screening is cost effective.³⁵ The specificity and NPV for the integrated ANC panel triple testing and standard of care were the same across the three infections that were screened for. Although it was not possible to determine the sensitivity and specificity for HBV, the performance of ANC panel for integrated testing had the same pattern as the standard of care among the negatives. The panel picked an equal number of negatives compared with the standard of care across the HIV, syphilis and HBV infections that were screened for.

Conclusion and recommendations

The integrated testing panel has the same efficacy as routine care for HIV, syphilis and HBV screening in ANC settings, as it produced results that were comparable with the standard of care. The panel can be used for simultaneous screening for HIV, syphilis and HBV with a single prick in POC settings.

We recommend studies on economic evaluation to show value for money using this test compared to the status quo and how the approach improves the quality of care. We further recommend a similar study with a larger sample size for HBV be carried out.

Acknowledgements

We would like to acknowledge the contributions of the Ministry of Health and Child Care Leadership Head Office, National Institute of Health Research Director, Mashonaland East Province Provincial Medical Directorate, Mudzi District Medical Officer, study team at participating sites and the patients who agreed to take part in the study in Zimbabwe. Africa Centres for Disease Control and Prevention (CDC) organised a Scientific Manuscript Writing Workshop with the Journal of Public Health in Africa, in Kigali, Rwanda, where a draft was prepared.

The authors reported that they received funding from Abbott Diagnostics which may be affected by the research reported in the enclosed publication. The author has disclosed those interests fully and has implemented an approved plan for managing any potential conflicts arising from their involvement. The terms of these funding arrangements have been reviewed and approved by the affiliated University in accordance with its policy on objectivity in research. The author, L.C., serves as an editorial board member of this journal. The peer review process for this submission was handled independently, and the author had no involvement in the editorial decision-making process for this article.

N.H., P.N. and M.J.C. conceived the study, performed the investigation and project administration. N.H., P.N., T.M. and M.J.C. designed the methodology. N.H., T.M. and M.J.C. used software for data collection and analysis. N.H. and L.C. were responsible for resources and funding acquisition for data collection, analysis and submission of article. P.N., L.C. and M.J.C. provided supervision during drafting of the article. All authors carried out the experiments and took part in data curation, formal analysis, writing-original draft, visualisation, validation, and writing-review & editing the article.

Funding for data collection activities was provided by Abbott Diagnostics. However, Abbott Diagnostics had no role in the development of the protocol, submission for ethical review, actual data collection, analysis, preparation of the manuscript and submission for publication.

Data inputs and analytic code available upon request to the corresponding author, N.H.

The views and opinions expressed in this article are those of the authors and are the product of professional research. The article does not necessarily reflect the official policy or position of any affiliated institution, funder, agency or the publisher. The authors are responsible for this article’s results, findings and content.

References

Appendix 1

FIGURE 1-A1: Zimbabwe national rapid testing algorithm for human immunodeficiency virus and syphilis in pregnant women.

Appendix 2

FIGURE 1-A2: Testing strategy for hepatitis B virus.

Appendix 3

Antenatal care panel standard operating procedure

1.0 Purpose

Antenatal care (ANC) Panel is an immunochromatography assay for the simultaneous detection of human immunodeficiency virus (HIV), syphilis, hepatitis B virus (HBV) plus malaria infection.

2.0 Principle

The panel is comprised of four strips (Determine™ HIV Early Detect, Determine™ HBsAg 2, Determine™ Syphilis TP plus NxTek^TM Eliminate Malaria PF) for the detection of HIV, HBV, syphilis and malaria.

The individual strips in the panel function according to the following principles:

2.1 Determine™ human immunodeficiency virus early detect

This test follows immunochromatography for qualitative detection of HIV-1 p24 antigen, and antibodies to HIV-1 and HIV-2 that are non-immunocomplexed (free). The sample mixes with anti-p24 antibodies that are biotinylated and selenium colloid-conjugates that are coated with recombinant antigens from HIV-1, HIV-2 and HIV-1 group O, synthetic HIV-2 proteins and anti-p24 mouse monoclonal antibody.

The mixture migrates along a solid phase to recombinant HIV-1/HIV-1 group O Ags and synthetic HIV-1/HIV-2 peptides that are immobilised at the antibody (Ab) window before reaching avidin immobilised at the antigen (Ag) window. If HIV-1 and/or HIV-2 Abs are present in the specimen, they bind to the selenium colloid-conjugates that are coated with recombinant Ags from HIV-1, HIV-2 and HIV-1 group O and synthetic HIV-2 proteins and to the immobilised recombinant HIV-1/HIV-1 group O Ags and synthetic HIV-1/HIV-2 proteins to form a single red line at the Ab window area.

When there are no HIV-1 and HIV-2 antibodies, the selenium colloid conjugates flow beyond the Ab window, resulting in no red line formation at the Ab window area.

If non-immunocomplexed HIV-1 p24 Ag is present in the specimen, it binds to both the biotinylated anti-p24 Abs and the selenium colloid conjugate which is coated with anti-p24 mouse monoclonal Ab. This complex is captured by the immobilised avidin, forming a red line at the Ag window area.

When there is no HIV-1 p24 Ag, the biotinylated anti-p24 Abs and selenium colloid conjugate move past the Ag window and no red line forms at the Ag window area.

A procedural control line is incorporated in the assay device, at the Control window, to show that the procedure has been carried out correctly.

2.2 Determine™ hepatitis B surface Ag 2

This test employs chromatography for qualitative detection of hepatitis B surface Ag (HBsAg). The sample is mixed with biotinylated anti-HBsAg mouse monoclonal antibodies and black particles that are coated with anti-HBsAg mouse monoclonal antibodies. This mixture then migrates laterally, along a solid phase towards immobilised avidin at the patient window. If HBsAg is present in the specimen, it binds to both the biotinylated anti-HBsAg mouse monoclonal antibodies and to black particles coated with anti-HBsAg mouse monoclonal antibodies. This complex is subsequently captured by the immobilised avidin, resulting in the appearance of a black line on the test strip.

If HBsAg is absent in the patient sample, the black particles migrate past the patient window and no black line is formed on the test strip. A procedural control line in the assay device at the Control window is incorporated to show that the procedure has been carried out correctly.

2.3 Determine™ syphilis TP

This test employs chromatography for qualitative detection of Abs to Triponema pallidum Ags. When the patient sample is added to the sample pad, it migrates through a conjugate pad, mixing with Triponema pallidum Ag-selenium colloid conjugate. The mixture continues to move through the solid phase to the immobilised Triponema pallidum Ags at the patient window. If Triponema pallidum Abs are present, in the sample, the Abs bind to the Triponema pallidum Ag-selenium colloid and the Triponema pallidum Ag at the patient window, forming a red line. If Triponema pallidum Abs are absent, the Triponema pallidum Ag-selenium colloid migrates past the patient window. No red line will appear at the patient window. A procedural control line in the assay device at the Control window is incorporated to show that the procedure has been carried out correctly.

3.0 Performance characteristics and measurement of uncertainty

4.0 Type of specimen

Capillary whole blood

5.0 Patient preparation

No special patient preparation is required. The patient just goes through the screening, enrolment and informed consent procedure as detailed in the protocol.

6.0 Type of container and additives

Not applicable. No sample is going to be collected for later use.

7.0 Specified equipment, reagents and supplies

7.2 Materials required but not provided

1. Timer

2. Sharps container

3. Protective gloves

4. Pen

8.0 Environmental and safety controls

• 8.1 All patient samples, reagents and controls should be treated as potentially infectious and appropriate precautions should be taken when handling them and on disposal.
• 8.2 Wear protective gloves when performing the test.
• 8.3 Disposable apparatus must be treated as biohazardous waste.
• 8.4 All kit components in the Panel pouch are for single use.
• 8.5 All sharps must be discarded in a sharps container.

9.0 Calibration procedures

Not applicable.

10.0 Specimen preparation

There is no special specimen preparation. Capillary whole blood is used for the testing using the ANC panel.

11.0 Testing procedure

• 11.1 Put on gloves.
• 11.2 Remove testing devices from the pouches.
• 11.3 Write participant identification number on each of the four testing devices.
• 11.4 Ask the participant to warm the fingers by rubbing to increase blood flow.
• 11.5 Select a fingerstick site off centre of the 3rd of the 4th fingertip.
• 11.6 Have the participant hold their hand so that the level of the finger is below the elbow to increase blood flow to the fingertips.
• 11.7 Wipe the fingertip with an alcohol swab and allow the alcohol to air dry.
• 11.8 Hold the finger still.
• 11.9 Place the lancet off centre of the fingertip.
• 11.10 Press the lancet firmly into the finger to puncture the skin.
• 11.11 Discard the used lancet into a sharps container immediately.
• 11.12 Gently wipe off the first drop of blood.
• 11.13 Carefully apply repeated pressure and release along the finger and hand except the puncture site. Avoid squeezing the finger.
• 11.14 Hold the EDTA Sample Collection Tube level and horizontal to the finger and place it in direct contact with blood drop. Do not press the capillary tube against the finger.
• 11.15 Fill the EDTA Sample Collection Tube completely.
• 11.16 Stop filling when the collection reservoir is full. At least 200 µL of blood has to be collected (up to the 2nd line).
• 11.17 Pull the capillary out of the collection tube and discard it in a sharps container.
• 11.18 Close the EDTA sample collection tube.
• 11.19 Turn the tube upside down 8–10 times to thoroughly mix the blood.
• 11.20 Place the tube back onto the work tray.

Running the determine™ human immunodeficiency virus early detect test

• 11.21 Remove the protective foil cover from the labelled testing device.
• 11.22 Open the EDTA sample collection tube.
• 11.23 Take the HIV capillary tube, hold it horizontally and touch the tip of the tube into the blood in the EDTA sample collection tube.
• 11.0 Let the Capillary Tube fill with blood
• 11.1 Hold the capillary tube in this position until the sample has reached between the two marked lines (50 µL) then remove it.
• 11.2 Close the EDTA sample collection tube.
• 11.3 Place the Capillary Tube onto the middle of the HIV test sample pad marked by the arrow symbol, then hold it in an upright vertical position.
• 11.4 Transfer the sample to the sample pad. Avoid lifting the capillary tube from the pad until complete transfer of the sample.
• 11.5 Discard the capillary tube in a sharps container.
• 11.6 Apply one drop of chase buffer to the sample pad immediately.

Running the determine™ syphilis TP test

• 11.7 Remove the protective foil cover from the syphilis test
• 11.8 Open the EDTA sample collection tube.
• 11.9 Take the syphilis capillary tube, hold it horizontally and touch the tip of the tube into the blood in the EDTA sample collection tube.
• 11.10 Let the capillary tube fill with blood.
• 11.11 Hold the capillary tube in this position until the sample has reached between the two marked lines (50 µL) then remove it.
• 11.12 Close the EDTA sample collection tube.
• 11.13 Place the capillary tube onto the middle of the Syphilis test sample pad marked by the arrow symbol, then hold it in an upright vertical position.
• 11.14 Transfer the sample to the sample pad. Avoid lifting the Capillary Tube from the pad until complete transfer of the sample.
• 11.15 Discard the capillary tube in a sharps container.
• 11.16 Apply one drop of chase buffer to the sample pad immediately.

Running the determine™ hepatitis B surface Ag 2 test

• 11.17 Remove the protective foil cover from the HBsAg test.
• 11.18 Open the EDTA sample collection tube.
• 11.19 Take the HBsAg capillary tube, hold it horizontally and touch the tip of the tube into the blood in the EDTA Sample Collection Tube.
• 11.20 Let the capillary tube fill with blood.
• 11.21 Hold the capillary tube in this position until the sample has reached between the two marked lines (50 µL) then remove it.
• 11.22 Close the EDTA sample collection tube.
• 11.23 Place the capillary tube onto the middle of the HBsAg test sample pad marked by the arrow symbol, then hold it in an upright vertical position.
• 11.24 Transfer the sample to the sample pad. Avoid lifting the capillary tube from the pad until complete transfer of the sample.
• 11.25 Discard the capillary tube in a sharps container.
• 11.26 Apply one drop of chase buffer to the sample pad immediately.

12.0. Interpretation of results

Interpretation of human immunodeficiency virus results

• 12.4 Non-reactive: One line in the control window (labelled ‘C’), and no line in the Ab and Ag windows of the strips (labelled ‘AG’ and ‘AB’).
• 12.5 Reactive: Any line on 1) the Ab window alone, 2) Ag window alone or 3) both the Ab and Ag windows simultaneously suggest infection with HIV.
• 12.6 Invalid: No line in the Control window, even if there are lines in the Ag and/or Ab windows.

Interpretation of the syphilis results

• 12.7 Negative: One line in the control window (labelled ‘C’), and no line in the patient window (labelled ‘Patient’).
• 12.8 Positive: Two lines in both the Control window (labelled ‘Control’) and patient window (labelled ‘Patient’) of the strip.
• 12.9 Invalid: No line in the Control window, even if there is a line in the patient window.

Interpretation of the hepatitis B surface Ag 2 results

• 12.10 Non-reactive: One line in the control window
• 12.11 Reactive: Two lines in both the Control window and patient window of the strip.
• 12.12 Invalid: No line in the Control window, even if there is a line in the patient window.

13.10 Quality control

Inhouse negative and positive controls are run before testing participants to determine if the devices are working properly.

14.0 Limitations

Determine™ human immunodeficiency virus early detection test

• 14.1 The test should only be used for human serum, plasma, venous blood and capillary blood. It should not be used for other body fluids or pooled samples.
• 14.2 The intensity of the AB and AG lines does not necessarily correlate to the Ab and Ag titer respectively in the sample.
• 14.3 A reactive result for Abs to HIV1/2 combined with a non-reactive result for HIV-1 p24 Ag does not preclude the possibility of acute HIV infection.
• 14.4 Both reactive Ag and Ab lines or Ag line only may be confirmed by a 4th generation EIA or NAT where the rapid diagnostic test (RDT) is indicated as a diagnostic aid. Ab reactivity only can be confirmed also by use of another HIV Ab IVD. If non-reactive in confirmatory testing, retest in 14 days following the date of the original test when the procedure is being followed for diagnostic purposes. Local algorithm is followed.

  There is no test that provides absolute assurance that a specimen does not have HIV-1 p24 Ag and/or Abs to HIV-1 and HIV-2 in low levels, such as at either very early stage or late stage of infection with HIV.

• 14.5 A non-reactive result for both Abs to HIV-1/2 and HIV-1 p24 Ag does not preclude the possibility of exposure to or infection with HIV-1/2 viruses.
• 14.6 The absence of AG line may occur when all p24 Ag is bound by Abs. After seroconversion, high levels of anti-p24 Abs will result in the Abs binding to the p24 Ags forming immunocomplexes. Determine™ HIV early detection test only detects non-immunocomplexed Ags, it does not detect immunocomplexed (bound) Ags.
• 14.7 Some individuals on antiretroviral therapy (ART) may produce false negative results by RDTs.
• 14.8 Samples containing anticoagulants besides EDTA may give incorrect results with Determine™ HIV early detection test.
• 14.9 The test cannot be used in neonates exposed to HIV.
• 14.10 Biotin treatment higher than 20mg/day may lead to decreased Ag line intensity.

Determine™ syphilis TP test

• 14.11 The test is designed to detect Abs to Treponema pallidum in human serum, plasma and whole blood. It should not be used for other body fluids or pooled samples.
• 14.12 The intensity of the patient line does not necessarily correlate to the titre of Ab in the sample.
• 14.13 No test provides absolute assurance that a sample does not contain low levels of Abs to Treponema pallidum such as those present at a very early stage of infection or low reactivity to Treponema pallidum Ags. Therefore, a negative result does not preclude the possibility of exposure to syphilis.
• 14.14 Biotin treatment higher than 200mg/day may lead to decreased control line intensity.

Determine™ hepatitis B surface Ag 2 test

• 14.15 The test is designed to detect HBsAg in human serum, plasma and whole blood. It should not be used for other body fluids or pooled samples.
• 14.16 The intensity of the patient line does not necessarily correlate to the titre of Ag in the sample.
• 14.17 There is no test that provides absolute assurance that a specimen has no low levels of HBsAgs such as those present during very early stage of infection.
• 14.18 If the result is non-reactive at 15 minutes and becomes reactive at 30 min, the participant has HBsAg that is very low, below the detection limit.
• 14.19 A non-reactive result does not rule out the possibility of exposure to hepatitis B or infection with the viruses.
• 14.20 Samples containing anticoagulants besides EDTA may give incorrect results with Determine™ HBsAg 2 test.
• 14.21 Biotin treatment higher than 5mg/day may lead to decreased participant line intensity.

15.0 RECORDS

16.0 Reference

• 1. Antenatal care panel product insert