M.Sc. (Chemistry) / With the increase in the number of new Human Immunodeficiency Virus (HIV) infection and mortality rate worldwide partly due to diagnostic drawbacks which gives false negatives during the window period, it is imperative to find an alternative method of detection. The need for prompt, bed-side and field applicable analytical devices for biomedical purposes cannot be over emphasized in our fast paced world today. As a contribution to meeting these challenges, this dissertation reports (i) the development of novel electrochemical DNA/RNA aptamer biosensor for HIV sensing and (ii) the development of other DNA sequence specific electrochemical biosensors. These biosensors were based on composite platforms of dendrimer and gold nanoparticles...

Submitted in fulfillment of the requirements for the Master of Health Sciences in Nursing, Durban University of Technology, Durban, South Africa, 2017. / Background Human immunodeficiency virus (HIV) testing, treatment and prevention programmes have been initiated and implemented, but nearly 19 million of the 35 million people globally who are living with HIV do not know they have it. A new and powerful movement called 90-90-90 has been set in motion where the target is that by the year 2020, 90% of all people should know their HIV status, 90% of those testing HIV positive should be commenced on lifelong antiretroviral treatment and 90% of the people receiving treatment should be virally suppressed. It is argued the new innovative HIV self-testing strategy can increase the uptake of HIV testing among key populations and the general public. Aim of the study The aim of the study was to assess health care users’ knowledge, attitudes and perceptions towards HIV self-testing at selected Gateway clinics in eThekwini District, KwaZulu-Natal. Methodology A quantitative, non-experimental descriptive design was used to determine knowledge, attitudes and perceptions of health care users at three selected Gateway clinics in eThekwini Health District. The researcher requested permission and was granted to conduct the study from all the relevant stakeholders. Human rights were protected and ethical considerations were adhered to throughout the research process. The convenience sample was 442 participants with a minimum of 98 and a maximum of 246 participants sampled from each of the three study sites. A survey questionnaire was used to collect data. Version 23 of SPSS was used to analyse data. Graphs and tables were used to represent frequencies. Inferential statistical were used to test whether any of the response options were selected significantly more or less than others Results Results of the study revealed that health care users had a reasonable knowledge of HIV self-testing and there were indications that they would use it if it was made freely available to the public and was properly regulated. Generally, there were indications that health care users had positive attitudes towards HIV self-testing. It was seen as a good idea as it can be performed in the privacy of one’s home and the person would be first to know about the results. Results also revealed that there could be more people who would know their HIV status and people could test more frequently. There were perceptions that there would be no difficulty in performing an HIV self-test. The lack of pre-test counselling, false negative results, possible coercion and sale of unregulated testing kits seemed to be issues of concern that require addressing if HIV self-testing is to be promulgated. / M

Paediatric human immunodeficiency virus (HIV) infection has become a major socio-economic health problem in recent years as the number of HIV-1 infected children steadily increases. The majority of these infants are infected through mother-to-child transmission, with the frequency of vertical transmission varying between 12,9% and 65%. In order to implement appropriate management and possible treatment of these infected neonates, it is essential to have reliable laboratory tests for the early diagnosis of an HIV infection. At the time that this study was initiated, the diagnosis of HIV-1 infection in the Groote Schuur Hospital Virology Laboratory depended almost exclusively on serological assays. Such assays are of limited value for infants under 18 months of age, as maternal lgG antibody to HIV-1 is transferred via the placenta and may persist in the baby for up to 18 months. Available lgG antibody tests do not distinguish reliably between passively acquired maternal antibody and that produced by the infant itself. A valuable method of establishing the presence of true infection is provided by the polymerase chain reaction (PCR) technique which allows the identification, and subsequent exponential amplification of low levels of specific viral nucleic acid using specific oligonucleotide primers. A major aim of this study was to develop and instigate a (PCR) assay for the early diagnosis of HIV infection in infected infants. This was successfully achieved by the adaptation and optimization of an existing standard PCR protocol to suit the specific needs of a routine diagnostic service. Preliminary requirements involved the selection of primers and probes and establishing optimal parameters for: ionic strength, Taq DNA polymerase concentration, primer concentration, deoxynucleotide triphosphate concentration, and hybridization conditions for most efficient functioning of the test. The devised method entailed the extraction of proviral DNA from peripheral blood mononuclear cells, amplification of HIV-1 specific sequences by PCR, and identification by Southern blot hybridization with digoxigenin (DIG)-labelled probes. Thereafter the efficacy of the assay was tested on 45 infants (under 15 months of age) all born to seropositive mothers and therefore at risk for HIV infection. Forty-two of these infants had antibodies to HIV-1 and the remaining 3 were seronegative. The latter 3 also tested negative for HIV proviral DNA when PCR was performed, using at least 2 different HIV-1 primer pairs and their respective DIG-labelled probes. However, 27 (64%) of the 42 seropositive infants were also HIV-PCR positive and the remaining 15 (36%) seropositive infants were negative for HIV proviral DNA. Positive PCR tests correlated well with clinical data indicative of active HIV-1 infection for the majority of infants in the neonatal period, although it could not provide proof of infection in newborn babies (less than 1 week of age). The development of an in-house PCR protocol specific for HIV-1 has not only provided a valuable diagnostic assay for neonatal infection, but has also given insight into the parameters required for high sensitivity and the stringent precautionary measures that need to be applied to avoid contamination problems. The second part of this study was devoted to DNA sequence analysis of cloned HIV isolates from an infected mother and her 3-month-old infant. Nucleotide sequence variation between isolates of HIV-1 has been well documented. Examination of the third variable region (particularly the V3- loop) in the env gene of HIV-1 of our mother-infant pair confirmed this variation and provided the first genetic epidemiological data of this nature in the local community. Proviral DNA from both mother and baby was amplified using V3-specific degenerate primers and cloned. Clones containing the insert DNA were 2 identified by colony-blot hybridization. Their nucleotide and amino acid sequences were analyzed by using various computer programs. The degree of similarity between variants from the mother and infant in this study differed to a large extent from previous studies. The virus population harboured by the mother displayed highly homogeneous V3 sequences (1,04% variation) compared to the isolates from her 3-month-old infant, which showed a higher degree (1,8%) of heterogeneity. Phylogenetic analysis of the different isolates from mother and infant demonstrated that an HIV-1 subtype C virus was the infectious agent. This classification was confirmed by the characteristic amino-acid sequence of the tetrapeptide motif of the V3 loop present in the isolates from both mother and infant as well as the absence of a potential N-linked glycosylation site proximal to the first cysteine of the V3 loop, which is characteristic of subtype C viruses. Based on the amino acids present at positions 306 and 320 of the V3 loop, it could also be concluded that isolates from both the mother and her baby were consistent with the non-syncytium inducing (NSI) phenotype of HIV-1, thus indicating that, contrary to popular belief, NSI variants can be responsible for initiating infection. Data obtained from these genetic investigations of variants involved in vertical transmission of HIV-1 can form a useful basis for future comparative studies.

Thesis (M. Tech.) - Central University of Technology, Free State, 2010 / Early detection of mother-to-child transfer of Human Immunodeficiency Virus (HIV-1) is of the utmost importance for monitoring the success of intervention strategies, as well as for optimal treatment of HIV-positive children. Serology can only be used confidently after 18 months, as remaining antibodies from the mother may give false positive results. This leaves only molecular methods for early detection of the virus; unfortunately, the technology is still too expensive for general use. The aim of this project was to develop and validate a cost-effective, fast, early detection method for HIV infection in infants. PCR was chosen as the developmental method, a technique that amplifies proviral sequences of HIV DNA, detecting HIV infection in peripheral blood mononuclear cells (PBMC) from infants of seropositive women during neonatal (age less than 28 days) and post-neonatal periods. A method based on the commercial Roche HIV-1 DNA assay was chosen for implementation on the Roche LightCycler instrument. The published primer set was used to detect both HIV-1 DNA and an internal control. The target DNA for use as internal control was constructed from the plasmid pBR322 so that an AT-rich part of the plasmid was flanked by the HIV-1 primer-binding sites. The resulting amplicon was cloned into a vector and multiplied in E. coli. Amplification of the plasmid by PCR in the Roche LightCycler in the presence of SYBR Green created an amplicon having a Tm different (81 ± 1°C ) from that of the HIV-1 amplicon (84 ± 1°C) so that post-amplification melting can be used to differentiate between HIV-1 and internal control. After construction of the internal control, the reaction conditions were optimised so that the internal control would amplify strongly only in the absence of HIV-1 target DNA. Then 50 previously tested patient samples were analysed using the assay developed here. Only half of the known positive samples came up positive in the assay, indicating that it is not sensitive enough for diagnostic use in its current form. Various ways of improving the sensitivity are suggested for further development of the assay as described here.

Submitted in fulfillment of requirements for the Degree of Masters in Technology: Nursing, Durban University of Technology. 2013 / This study investigated the factors that impact on the utilisation of voluntary HIV counselling and testing (VCT) service at a wellness centre in a higher education institution. Purpose: The purpose of the study was to identify factors that impact on the utilisation of VCT service in a higher education institution. Methodology: A quantitative descriptive survey research design was used to describe the phenomenon and to establish relationships between variables. Participants were selected randomly in order to obtain a broad representative sample in three strata. A formal structured close-ended questionnaire was used to collect data. The questionnaire was designed to focus on variables such as demographic data, environmental factors, enabling factors, predisposing and behavioural factors. Findings: No significant relationship was identified between knowledge of HIV, AIDS and VCT and high school attended in rural, township and urban areas. There was no significant relationship between environmental factors and utilisation of VCT. There was no association between predisposing factors such as risk of VCT and perceived benefits of VCT utilisation. There was no relationship between knowledge and consistency of condom use and utilisation of VCT. The majority of respondents stated that the VCT service was not user friendly and the attitude of the staff toward students was poor. There was no significant relationship between accessibility of VCT service and utilisation of VCT. There was a lack of utilisation of campus VCT services and the majority of respondents utilised other services. Conclusion: The study showed that respondents who knew about availability of VCT were likely to have used the facility. Respondents who showed positive / Durban University of Technology

Thesis (MPhil)--University of Stellenbosch, 2007. / ENGLISH ABSTRACT: Voluntary Counseling and Testing (VCT) is considered an important component of the South African government’s response to the HIV/AIDS pandemic. Despite of this, the numbers of South Africans who have accessed VCT remains low. Pregnant women in antenatal clinics remain the primary recipients of HIV testing since it is routinely offered to them at antenatal clinics. Figures from antenatal clinics constitute the most important source of HIV/AIDS statistics in South Africa. Targeting women at antenatal clinics perpetuate the stereotypes that women are solely responsible for reproductive health issues or, that they are to be blame for the mounting HIV infections. These beliefs exacerbate the culminating violence against women. According to observation, active involvement of male youth in reproductive health at the Rosedale Clinic situated in Uitenhage, a town in the Eastern Cape, appears to be very limited. This might be as a result of traditional beliefs declaring reproductive health a primarily female domain. Active involvement of males in reproductive health is considered to have positive health outcomes both for themselves, and their partners. The purpose of this study is thus to encourage males served by the Rosedale clinic to become more actively involved in one aspect of reproductive health, namely voluntary counseling and testing (VCT). The study aims to find out which factors constitute barriers and enablers to VCT acceptance by male youth. Semi-structured, face-to- face interviews were conducted in September 2006 with eighteen males, 18 to 25 years, to determine from their perspective what constitute barriers and enablers to HIV test acceptance by males. These participants were recruited from two well-known rugby clubs in Uitenhage, located in the Eastern Cape. In addition, a focus group discussion was conducted with four males recruited at a graduate development programme hosted in Uitenhage in order to explore common themes emerging from interviews and the discussion. A pervasive, psychologically rooted fear; and possible low perception of risk emerge from this study as main barriers to HIV testing. Negative perceptions regarding service delivery in the public health sector constitute another barrier to HIV testing. The possibility of teasing, a common phenomenon amongst rugby players but not exclusive to them, exacerbates peer pressure thus possibly contributing to low levels of HIV test acceptance among these males. Increased HIV test acceptance is associated with increased opportunities of exposure to HIV testing opportunities, for example at awareness campaigns. Access to medical aid and income is also associated with an increased likelihood of HIV testing since most males prefer accessing HIV testing services in the private sector. Convenience and ease of access to HIV testing services are important when these males consider going for HIV testing. / AFRIKAANSE OPSOMMING: Vrywillige Berading en MIV Toetsing, een aspek van reproduktiewe gesondheid, word beskou as ‘n belangrike komponent van die Suid-Afrikaanse regering se strategie teen MIV/VIGS. Ongeag van hierdie feit is ‘n groot aantal Suid-Afrikaners nog nie getoets vir MIV nie. Swanger vrouens word op ‘n daaglikse basis hierdie toets aangebied as deel van roetine ondersoeke. Syfers vanaf pre-natale klinieke vorm die hoofbron van Suid- Afrika se HIV/VIGS statistieke. Die feit dat swanger vrouens die hoofteikengroep is vir MIV toetsing dra by tot stereotipes wat daartoe lei dat vrouens primêr verantwoordelik is vir die behoud van reproduktiewe gesondheid; en dat hulle beskou word as die hoofverdagtes in die toenemende oordrag van MIV. Laasgenoemde beskuldiging vererger geweld teen vrouens. Volgens waarneming by Rosedale publieke kliniek geleë in Uitenhage, ‘n dorp in die Oos-kaap, is veral jeugdige mans se betrokkenheid in reproduktiewe gesondheid uiters minimaal. Dit kan moontlik wees omdat reproduktiewe gesondheid tradisioneel as ‘n vroulike domein beskou word. Aktiewe betrokkenheid van beide mans en vrouens in reproduktiewe gesondheid het positiewe gesondheidsuitkomste vir beide partye tot gevolg. Die doel van hierdie studie is dus om jongmans wie deur die Rosedale kliniek bedien word aan te moedig om meer sigbaar te word in reproduktiewe gesondheidskwessies; met die oog op een aspek daarvan naamlik, vrywillige berading en MIV toetsing. Die studie poog dus om uit te vind watter struikelblokke jeugdige mans weerhou van MIV toetsing en; watter faktore mans aanmoedig om vir die toets te gaan. Semi-gestruktureerde, aangesig-tot- aangesig onderhoude is in September 2006 met 18 mans, vanaf die ouderdomme 18 tot 25 jaar, gevoer. Hierdie mans was tydens die studie rugbyspelers vanuit twee welbekende rugbyklubs op die dorp. ‘n Addisionele fokus groep is verder gevoer met manlike studente wie ten tye van die studie aan ‘n ontwikkelingsprogram vir gradueerders deelgeneem het op die dorp. Hierdie fokusgroep is geloods om soortgelyke temas vanuit die onderhoude en die bespreking te ondersoek. Die studie toon dat ‘n diepgewortelde, sielkundige vrees; en moontlike lae bewustheid van persoonlike risiko die twee vernaamste struikelblokke is vir MIV toetsing. Negatiewe sienings rakende die publieke gesondheidssektor kan moontlik bydrae tot weerstand teen MIV toetsingsdienste. Tergery, ‘n bekende verskynsel onder rugbymans, maar nie slegs beperk tot diè groep nie, dra by tot groepsdruk en weerhou moontlik menige mans van MIV toetsing. Diegene in die studie wie alreeds vir MIV getoets is geniet toenemende blootstelling aan MIV toetsingsgeleenthede; bv tydens bewusmakingsveldtogte geloods òf by die werk òf tersiêre instellings. Toegang tot ‘n mediese fonds; en ‘n inkomste is moontlike bepalende faktore sienende dat meeste respondente gesondheidsdienste in die privaatsektor verkies. Gerieflike toegang tot MIV toetsingsdienste word deur sommige mans as belangrike beskou wanneer hulle MIV toetsing oorweeg.

This thesis examines HIV disease natural history and response to antiretroviral treatment (ART) in patients from The TREAT Asia HIV Observational Database (TAHOD), a mulitcentre, prospective observational cohort of HIV-infected patients from countries in the Asia-Pacific region. By September 2005, 2979 patients have been recruited to TAHOD from 15 participating sites. The majority were male (73%), median age 37 years. Chinese (37%), Thai (25%) and Indian (17%) were the main ethnicities. Most patients reported HIV infection through heterosexual (59%) and homosexual contact (23%); 5% injecting drug use. At baseline, 41% of patients were diagnosed with AIDS defining illness, and 77% were being treated with highly active antiretroviral therapy (HAART). Baseline and retrospective data suggest that the overall response to HAART in TAHOD is similar to that seen in western cohorts, with mean CD4 count increase of 115 cells/μL and 69% achieving a viral load less than 400 copies/mL six-month after HAART initiation. Baseline CD4 count was the strongest predictor of short-term disease progression. Prognostic models based on routine clinical data and haemoglobin gave a good estimation of disease progression. The rate of new AIDS defining illness was 26% in the first 90 days after HAART initiation, which may partly be due to immune reconstitution syndrome occurring shortly after treatment. The most frequently used first-line ART combination was stavudine/lamivudine/nevirapine. Approximately 22% of patients receiving this treatment changed or stopped at least one drug in the first year, with adverse effect (including lipodystrophy, hepatitis, rash and peripheral neuropathy) the major reasons. The rates of discontinuation of efavirenz or nevirapine as part of HAART were similar (16 vs. 20/100 person years). Older age and positive HCV antibody were associated with an elevated liver function (ALT) test. Both prevalence of HBV and HCV coinfection with HIV were approximately 10%. The impact of hepatitis coinfection on immunological and virological responses to ART and HIV disease progression was not statistically significant. Both HBV and HCV remained independently associated with elevated ALT in the multivariate models. The overall HIV disease progression and response to ART in TAHOD patients were similar to those seen in the western countries.

HIV testing is required for life assurance applications. A written information document distributed at blood collection (venisection) serves as pre-test preparation. This study reviewed the adequacy of the document and explored possible alternative arrangements, by means of three research phases conducted at the point of venisection. Phase 1 used a specifically designed questionnaire which included a demographic section and questions assessing the applicant's appraisal of being adequately prepared, and their understanding and experience of testing. Constructed measures established their state of anxiety at testing and their range of information about HIV/AIDS. Phase 2 consisted of a counselling intervention, followed by the questionnaire used in Phase 1. Phase 3 consisted of semi-structured interviews with nursing personnel and insurance brokers. The phase 1 results indicated that the majority of applicants knew they were having an HIV test, did not feel coerced, had a moderate level of information about HIV and were not overly anxious at testing. The level of information about HIV/AIDS showed a significant correlation with their level of education, and the information document emerged as inadequate preparation. Answers given in Phase 2 differed qualitatively from those in Phase 1. Greater consideration of the impact of a positive result was shown, with increased concern about the implications for other people and anticipated acceptance of a positive result emerged in Phase 2. The personnel interviewed for Phase 3 indicated that they felt ill equipped to offer pre-test preparation. / Thesis (M.A.)-University of Natal, Pietermaritzburg, 2001.

This thesis examines HIV disease natural history and response to antiretroviral treatment (ART) in patients from The TREAT Asia HIV Observational Database (TAHOD), a mulitcentre, prospective observational cohort of HIV-infected patients from countries in the Asia-Pacific region. By September 2005, 2979 patients have been recruited to TAHOD from 15 participating sites. The majority were male (73%), median age 37 years. Chinese (37%), Thai (25%) and Indian (17%) were the main ethnicities. Most patients reported HIV infection through heterosexual (59%) and homosexual contact (23%); 5% injecting drug use. At baseline, 41% of patients were diagnosed with AIDS defining illness, and 77% were being treated with highly active antiretroviral therapy (HAART). Baseline and retrospective data suggest that the overall response to HAART in TAHOD is similar to that seen in western cohorts, with mean CD4 count increase of 115 cells/μL and 69% achieving a viral load less than 400 copies/mL six-month after HAART initiation. Baseline CD4 count was the strongest predictor of short-term disease progression. Prognostic models based on routine clinical data and haemoglobin gave a good estimation of disease progression. The rate of new AIDS defining illness was 26% in the first 90 days after HAART initiation, which may partly be due to immune reconstitution syndrome occurring shortly after treatment. The most frequently used first-line ART combination was stavudine/lamivudine/nevirapine. Approximately 22% of patients receiving this treatment changed or stopped at least one drug in the first year, with adverse effect (including lipodystrophy, hepatitis, rash and peripheral neuropathy) the major reasons. The rates of discontinuation of efavirenz or nevirapine as part of HAART were similar (16 vs. 20/100 person years). Older age and positive HCV antibody were associated with an elevated liver function (ALT) test. Both prevalence of HBV and HCV coinfection with HIV were approximately 10%. The impact of hepatitis coinfection on immunological and virological responses to ART and HIV disease progression was not statistically significant. Both HBV and HCV remained independently associated with elevated ALT in the multivariate models. The overall HIV disease progression and response to ART in TAHOD patients were similar to those seen in the western countries.

An HIV-positive diagnosis can be an overwhelming and traumatic experience. This study explores the experiences of receiving an HIV-positive diagnosis. Employing an Interpretative Phenomenological Analysis (IPA), a sample of the initial posts from threads on an online forum, was collected and explored, in order to determine the dominant themes from the experiences expressed in the posts, as well as the support sought from the forum. The online forum was accessed as an unobtrusive observer, and posts from January to December 2015 were explored. The online forum provides a platform for disclosure following an HIV-positive diagnosis, when anxiety and fear of stigma can impact on an individual’s ability to disclose to their social support network of family and friends. The experiences expressed on the online forum reflect the emotional, mental and physical impact of an HIV-positive diagnosis on an individual. The findings in this study reflected themes of shock, guilt and hopelessness, and concerns and fears regarding disclosure and stigma associated with HIV, as well as the importance of social support for the coping mechanisms of individuals after receiving an HIV-positive diagnosis. This research demonstrates the importance of Internet accessibility for information and support for chronic illnesses, such as HIV, and the role of the online forum platform for providing a safe environment for individuals recently diagnosed HIVpositive.