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Background We identified a HIV-positive cohort in virologic failure (VF) who re-suppressed without drug switch. We characterized their drug resistance mutations (DRM) and adherence profiles to learn how to better manage HIV drug resistance. A retrospective cohort study utilizing clinical data and stored samples. Patients received ART at three Nigerian treatment centres. Plasma samples stored when they were in VF were genotyped. Result Of 126 patients with samples available, 57 were successfully genotyped. From ART initiation, the proportion of patients with adherence ≥90% increased steadily from 54% at first high viral load (VL) to 67% at confirmed VF, and 81% at time of re-suppressed VL. Sixteen (28%) patients had at least one DRM. Forty-six (81%) patients had full susceptibility to the three drugs in their first-line (1 L) regimen. Thirteen (23%) were resistant to at least one antiretroviral drug but three were resistant to drugs not used in Nigeria. Ten patients had resistance to their 1 L drug(s) and six were fully susceptible to the three drugs in the recommended second-line regimen. Conclusion This cohort had little drug resistance mutations. We conclude that if adherence is not assured, patients could exhibit virologic failure without having developed mutations associated with drug resistance.

Nigeria has an emerging problem with multidrug-resistant tuberculosis (MDR-TB). Whole-genome sequencing was used to understand the epidemiology of tuberculosis and genetics of multi-drug resistance among patients from two tertiary referral centers in Southwest Nigeria. In line with previous molecular epidemiology studies, most isolates of Mycobacterium tuberculosis from this dataset belonged to the Cameroon clade within the Euro-American lineage. Phylogenetic analysis showed this clade was undergoing clonal expansion in this region, and suggests that it was involved in community transmission of sensitive and multidrug-resistant tuberculosis. Five patients enrolled for retreatment were infected with pre-extensively drug resistant (pre-XDR) due to fluoroquinolone resistance in isolates from the Cameroon clade. In all five cases resistance was conferred through a mutation in the gyrA gene. In some patients, genomic changes occurred in bacterial isolates during the course of treatment that potentially led to decreased drug susceptibility. We conclude that inter-patient transmission of resistant isolates, principally from the Cameroon clade, contributes to the spread of MDR-TB in this setting, underscoring the urgent need to curb the spread of multi-drug resistance in this region.

BackgroundAlthough there are a number of studies comparing the currently recommended preferred and alternative first-line (1L) antiretroviral therapy (ART) regimens on clinical outcomes, there are limited data examining the impact of 1L regimen choice and duration of virologic failure (VF) on accumulation of drug resistance mutations (DRM). The patterns of DRM from patients failing zidovudine (AZT)-containing versus tenofovir (TDF)-containing ART were assessed to evaluate the predicted susceptibility to second-line (2L) nucleoside reverse-transcriptase inhibitor (NRTI) backbone options in the context of an ongoing programmatic setting that uses viral load (VL) monitoring.MethodsPaired samples from Nigerian ART patients who experienced VF and switched to 2L ART were retrospectively identified. For each sample, the human immunodeficiency virus (HIV)-1 polymerase gene was sequenced at 2 time points, and DRM was analyzed using Stanford University’s HIVdb program.ResultsSequences were generated for 191 patients. At time of 2L switch, 28.2% of patients on AZT-containing regimens developed resistance to TDF, whereas only 6.8% of patients on TDF-containing 1L had mutations compromising susceptibility to AZT. In a stratified evaluation, patients with 0–6 months between tested VL samples had no difference in proportion compromised to 2L, whereas those with >6 months between samples had a statistically significant difference in proportion with compromised 2L NRTI. In multivariate analyses, patients on 1L AZT had 9.90 times higher odds of having a compromised 2L NRTI option than patients on 1L TDF.ConclusionsIn the context of constrained resources, where VL monitoring is limited, we present further evidence to support use of TDF as the preferred 1L NRTI because it allows for preservation of the recommended 2L NRTI option.

BACKGROUND: Drug-resistant tuberculosis (TB) is a global public health problem. Adequate management requires baseline drug-resistance prevalence data. In West Africa, due to a poor laboratory infrastructure and inadequate capacity, such data are scarce. Therefore, the true extent of drug-resistant TB was hitherto undetermined. In 2008, a new research network, the West African Network of Excellence for Tuberculosis, AIDS and Malaria (WANETAM), was founded, comprising nine study sites from eight West African countries (Burkina Faso, The Gambia, Ghana, Guinea-Bissau, Mali, Nigeria, Senegal and Togo). The goal was to establish Good Clinical Laboratory Practice (GCLP) principles and build capacity in standardised smear microscopy and mycobacterial culture across partnering laboratories to generate the first comprehensive West African drug-resistance data. METHODS: Following GCLP and laboratory training sessions, TB isolates were collected at sentinel referral sites between 2009-2013 and tested for first- and second-line drug resistance. RESULTS: From the analysis of 974 isolates, an unexpectedly high prevalence of multi-drug-resistant (MDR) strains was found in new (6 %) and retreatment patients (35 %) across all sentinel sites, with the highest prevalence amongst retreatment patients in Bamako, Mali (59 %) and the two Nigerian sites in Ibadan and Lagos (39 % and 66 %). In Lagos, MDR is already spreading actively amongst 32 % of new patients. Pre-extensively drug-resistant (pre-XDR) isolates are present in all sites, with Ghana showing the highest proportion (35 % of MDR). In Ghana and Togo, pre-XDR isolates are circulating amongst new patients. CONCLUSIONS: West African drug-resistance prevalence poses a previously underestimated, yet serious public health threat, and our estimates obtained differ significantly from previous World Health Organisation (WHO) estimates. Therefore, our data are reshaping current concepts and are essential in informing WHO and public health strategists to implement urgently needed surveillance and control interventions in West Africa.

Aim The study is aimed at assessing the knowledge, perception and acceptability of microbicides among non-health care workers in Lagos, Nigeria.

This study focuses on the identification of aetiological agents of vaginitis in Nigerian women. Study subjects are drawn from patients presenting with lower abdominal pain, vaginal discharge and itching at the gynaecology clinic of Lagos University Teaching Hospital and at the Clinical Centre of the Nigerian Institute of Medical Research, Yaba, Lagos, between January 2001 and July 2002. A total of 250 patients gave informed consent to participate in the study. The patients also had pre- and post-test human immunodeficiency virus (HIV) counselling. Each patient completed a questionnaire in order to provide biographical data, past clinical history and socio-economic background information. A cervical swab (CS) and a high-vaginal swab (HVS) were obtained from each patient. Swab samples were examined for pH and under light microscopy by Gram's stain and as wet preparations in 10% potassium hydroxide. Subsequently, samples were cultured on appropriate media at optimal conditions and a drug sensitivity profile for all isolates was determined by standard methods. Blood samples were screened and confirmed for HIV antibodies. Bacterial, fungal and parasitic pathogens were identified or isolated in samples from 241 (96.4%) of the women. Bacterial agents (Neisseria, Streptococcus and Staphylococcus species) were predominant in 128 (51.2%) patients, followed by fungi in 108 (43.2%) and parasites (Trichomonas vaginalis) in five (2.0%). Sensitivity to ciprofloxacin was seen in 40% of Staphylococcus species and in 90% of Neisseria species. Positive HIV serology was seen in 25 (10%) of the 250 women studied, 20 (80%) of which had concurrent microbial infections. Overall, a broad spectrum of microbial agents were shown to be responsible for vaginitis in the group of patients studied.

A diverse array of non-subtype B HIV-1 viruses circulates in Africa and dominates the global pandemic. It is important to understand how drug resistance mutations in non-B subtypes may develop differently from the patterns described in subtype B. HIV-1 reverse transcriptase and protease sequences from 338 patients with treatment failure to first-line ART regimens were evaluated. Multivariate logistic regression was used to examine the effect of subtype on each mutation controlling for regimen, time on therapy, and total mutations. The distribution of HIV-1 subtypes included CRF02_AG (45.0%), G (37.9%), CRF06_cpx (4.4%), A (3.6%), and other subtypes or recombinant sequences (9.2%). The most common NRTI mutations were M184V (89.1%) and thymidine analog mutations (TAMs). The most common NNRTI mutations were Y181C (49.7%), K103N (36.4%), G190A (26.3%), and A98G (19.5%). Multivariate analysis showed that CRF02_AG was less likely to have the M41L mutation compared to other subtypes [adjusted odds ratio (AOR) = 0.35; p = 0.022]. Subtype A patients showed a 42.5-fold increased risk (AOR = 42.5, p = 0.001) for the L210W mutation. Among NNRTI mutations, subtype G patients had an increased risk for A98G (AOR = 2.40, p = 0.036) and V106I (AOR = 6.15, p = 0.010), whereas subtype CRF02_AG patients had an increased risk for V90I (AOR = 3.16; p = 0.003) and a decreased risk for A98G (AOR = 0.48, p = 0.019). Five RT mutations were found to vary significantly between different non-B West African subtypes. Further study to understand the clinical impact of subtype-specific diversity on drug resistance will be critically important to the continued success of ART scale-up in resource-limited settings.

Population studies showed that there are differences in T-lymphocytes subpopulation of normal children in different regions, and reference values in an area might be different from another. This study compared the values in our population with CDC and WHO reference values. Blood samples from 279 healthy, HIV-negative children<12 years of age were analysed for complete blood count, CD3+, CD4+, CD8+ counts and percentages. Except for CD8%, mean values for all parameters measured significantly decreased with age. CD4+ counts were higher in females than males,P<.05. Using the WHO criteria, 15.9% of subjects had low total lymphocyte count and 20.6% had low CD4 count. Children<3 years had median CD4% lower than WHO normal values. Our median CD4+ counts correlated with CDC values. Values used by WHO in infants are higher than ours. We suggest that our children be assessed using CDC reference values which correlate with ours.

The gag p6 region of HIV-1 has various nonsubstitutionary mutations, including insertions, duplications, deletions, and premature stop codons. Studies have linked gag p6 mutations to reduced susceptibility to antiretroviral therapy in HIV-1 subtype B. This study examined the relationship between antiretroviral therapy and gag p6 diversity in HIV-1 CRF02_AG and subtype G. p6 data were generated for secondary analyses following Viroseq genotyping of pol gene sequences in plasma samples from HIV-1-infected Nigerians on reverse transcriptase inhibitor therapy, with virologic failure (repeat VL2000 copies/ml). p6 sequence chromatograms were available for 40 CRF02_AG and 43 subtype G-infected individuals. Subjects who had not received their supply of antiretroviral drugs for at least 2 months prior to the plasma sampling were classified as nonadherent. p6 sequences from therapy-adherent individuals had more nonsubstitutionary mutations than sequences from drug-naive individuals (p = 0.0005). The P5L/T mutation was inversely correlated with the presence of K27Q/N in p6, with each mutation being more prominent in subtype G and CRF02_AG, respectively. The data also suggested that P5L/T may be a compensatory mutation for the loss of an essential phosphorylation site in p6. In addition, there was an inverse association between P5L/T mutations in p6 and thymidine analog mutations in reverse transcriptase (p = 0.0001), and drug nonadherence was associated with an 8-fold lower risk of having a nonsubstitutionary mutation in p6 (95% CI = 1.27-52.57). Our data suggest that antiretroviral therapy influences gag p6 diversity, but further studies are needed to clarify these observations.

The study reported in this paper was carried out in the Northwestern and Southwestern regions of Nigeria, between October 1996 and April 1998. The study examined the possible contributory role of living conditions in the development of acute necrotizing gingivitis (ANG) or noma from oral lesions. Questionnaire data obtained from 42 fresh noma cases seen in the Northwest and four fresh cases seen in the Southwest were examined. In addition 46 cases of advanced ANG from the Southwest were included. The main focus was to compare some of the environmental living conditions of cases with advanced ANG and those with noma in these regions. All the noma and ANG cases were seen in children aged 2-12 years. The level of good oral hygiene practices and general environmental living conditions were significantly higher in the Southwest than in the Northwest. Data also showed that living in close proximity with livestock was significantly higher in the Northwest than in the Southwest (P < 0.05). The environmental living conditions of children in the Northwest were further compounded by poor sanitary faecal disposal practices as well as minimal access to potable water. The overall data indicated that living in substandard accommodations, exposure to debilitating childhood diseases, living in close proximity to livestock, poor oral hygiene, limited access to potable water and poor sanitary disposal of human and animal faecal waste could have put the children in the Northwest at higher risk for noma than the children in the Southwest. These could have been responsible for the higher prevalence of noma in the Northwest than in the Southwest.

Background: In order to scale up access to HIV counselling and testing in Nigeria, an HIV diagnostic algorithm based on rapid testing was adopted. However, there was the need to further evaluate the testing strategy in order to better assess its performance, because of the potential for false positivity. Objectives: The objective of this study was to compare positive HIV test results obtained from the approved rapid testing algorithm with results from western blot tests performed on samples from the same patient. Methodology: A retrospective review was conducted of HIV screening and confirmatory results for patients seen between 2007 and 2008. Rapid test and western blot results were extracted and compared for concordance. Discordant results were further reviewed using a combination of HIV-1 RNA viral load and CD4+ cell count test results and clinical presentation from medical records. Results: Analysis of 2228 western blot results showed that 98.3% ( n = 2191) were positive for HIV-1, 0.4% ( n = 8) were positive for HIV-2 and 0.3% ( n = 7) were dual infections (positive for both HIV-1 and HIV-2); 0.6% ( n = 13) were indeterminate and 0.4% ( n = 9) were negative. Further investigation of the 13 indeterminate results showed nine to be HIV-1 positive and four to be HIV-negative, for a total of 13 negative results. The positive predictive value of the HIV counselling and testing algorithm was 99.4%. Conclusion: Using the rapid testing algorithm alone, false positives were detected. Therefore, effective measures such as training and retraining of staff should be prioritised in order to minimise false-positive diagnoses and the associated potential for long-term psychological and financial impact on the patients.

The study was designed to examine the knowledge and perception of HIV positive persons about the antiretroviral therapy (ART) program and to determine their ability to pay for ART and the treatment of other opportunistic infections in Nigeria. This is aimed at identifying factors that may impede effective delivery and utilization of ART in the country. One hundred and twenty-five HIV positive persons seeking ART at the Nigerian Institute of Medical Research (NIMR) clinic, Lagos, were studied using questionnaires. Respondents' average monthly income was N11,253.00 (US$90.00). Almost 26% (25.6%) were unwilling to seek ART at the nearest hospital because of fear of stigmatization. While 9% wanted the therapy for free, the majority was willing to pay N500.00 (US$4.00) per month. The average affordable price based on the subjects' assessment was N905.00 (US$7.24), while the median was N500.00 (US$4.00) per month. Eighty-eight percent believed ART would prolong their lives. The ART drugs need to be affordable and building on the positive perceptions of ART is imperative.

The purpose of this study was to determine the bacterial diversity in advanced noma lesions using culture-independent molecular methods. 16S ribosomal DNA bacterial genes from DNA isolated from advanced noma lesions of four Nigerian children were PCR amplified with universally conserved primers and spirochetal selective primers and cloned into Escherichia coli . Partial 16S rRNA sequences of approximately 500 bases from 212 cloned inserts were used initially to determine species identity or closest relatives by comparison with sequences of known species or phylotypes. Nearly complete sequences of approximately 1,500 bases were obtained for most of the potentially novel species. A total of 67 bacterial species or phylotypes were detected, 25 of which have not yet been grown in vitro. Nineteen of the species or phylotypes, including Propionibacterium acnes , Staphylococcus spp., and the opportunistic pathogens Stenotrophomonas maltophilia and Ochrobactrum anthropi were detected in more than one subject. Other known species that were detected included Achromobacter spp., Afipia spp., Brevundimonas diminuta , Capnocytophaga spp., Cardiobacterium sp., Eikenella corrodens , Fusobacterium spp., Gemella haemoylsans , and Neisseria spp. Phylotypes that were unique to noma infections included those in the genera Eubacterium , Flavobacterium , Kocuria , Microbacterium , and Porphyromonas and the related Streptococcus salivarius and genera Sphingomonas and Treponema . Since advanced noma lesions are infections open to the environment, it was not surprising to detect species not commonly associated with the oral cavity, e.g., from soil. Several species previously implicated as putative pathogens of noma, such as spirochetes and Fusobacterium spp., were detected in at least one subject. However, due to the limited number of available noma subjects, it was not possible at this time to associate specific species with the disease.

Background Multidrug-resistant tuberculosis (MDR-TB) remains a clear threat to TB control. There is a paucity of data on DR-TB for many countries especially in sub-Saharan Africa. The study was undertaken to measure the prevalence of DR-TB, including MDR-TB, from West Africa. Methods Mycobacterial isolates were obtained from consecutive new and previously treated TB patients from Burkina Faso, Ghana, Guinea-Bissau, Mali, Nigeria, Senegal, The Gambia and Togo from December 2012 to December 2014. Phenotypic drug susceptibility testing to first- and second-line anti-TB drugs was performed using BACTEC MGIT 960 system. Results Viable isolates from a total of 44% (416/950) new and 56% (534/950) previously treated TB patients were included. HIV results were available for 599 (63%) with estimated HIV-TB co-infection of 21% (95% CI: 18.2−24.9%). Pooled estimate of any DR-TB prevalence among new TB patients was 20% (95% CI: 16.4−24.4%) while for MDR-TB this was 6% (95% CI: 4.1−9.0%). Among previously treated TB patients, these were 53% (95% CI: 48.3−56.9%) and 34% (95% CI: 30.1−38.3%), respectively. Significant factor for the development of MDR-TB was the history of previous anti-TB treatment (Crude OR=0.13; 95% CI: 0.08−0.20; p= Mono-resistance was detected in 12% (95% CI: 10.2−14.5%) with the highest resistance to streptomycin 6% (95% CI: 4.8−7.9%). Pooled estimate of pre-XDR-TB prevalence rate among MDR-TB patients was 21% (95% CI: 15.2−26.9%). Estimated resistance to ofloxacin, kanamycin, capreomycin and kanamycin and capreomycin were 7% (95% CI: 3.5−10.9%), 2% (95% CI: 0.6−5.1%), 9% (95% CI: 5.8−14.5%), and 3% (95% CI: 0.8−5.8%), respectively. Conclusions The reported prevalence of MDR-TB and pre-XDR-TB are high compared to WHO estimates. Resistance to streptomycin may indicate a high risk of failure for the WHO standard regimen. MDR-TB patients with resistance to either the fluoroquinolone or injectables may have suboptimal response; thus the need for continuous surveillance of TB resistance.

A previous study demonstrated the presence and possible involvement of Fusobacterium necrophorum in the pathogenesis of noma lesions of children living in agricultural and herding villages in northwestern Nigeria. In order to determine if F. necrophorum was part of the oral flora of malnourished children with no noma lesions, a study of the fusobacteria present in the oral cavities of 30 children, 2-6 years of age in Sokoto State, was undertaken. Swabs taken of the oral cavity were cultured on selective fusobacteria medium using conventional anaerobic microbiological techniques. F. nucleatum was recovered from each child and F. necrophorum was isolated from the oral cavity of only one child. The presence of F. nucleatum and the lack of F. necrophorum, except in one case, suggests that the latter is not normal flora in the children at risk for noma. F. necrophorum, a putative trigger organism for noma may gain a foothold only when certain staging conditions (i.e., lowered host resistance and/or oral lesion) are present.

Tuberculosis (TB) is the most common opportunistic infection in human immunodeficiency virus (HIV)-infected patients and the emergence of drug-resistant tuberculosis (DR-TB) is a growing problem in resource-limited settings. Adequate infrastructure for testing drug sensitivity and sufficient evidence of first-line resistance are currently unavailable in Nigeria. We collected sputum samples from HIV-infected patients enrolled in the Harvard PEPFAR/APIN Plus program over 12 months at two PEPFAR antiretroviral therapy (ART) clinics in the southwest and north central regions in Nigeria. Smear-positive sputum samples were submitted for GenoType MTBDRplus testing ( n = 415); mutations were confirmed through sequencing. Our results show high rates of DR-TB in Nigerian HIV-infected individuals (7.0% for rifampin [RIF] and 9.3% for RIF or isoniazid [INH]). Total RIF resistance indicative of MDR-TB in treatment-naive patients was 5.52%, far exceeding the World Health Organization predictions (0 to 4.3%). RIF resistance was found in 6/213 (2.8%) cases, INH resistance was found in 3/215 (1.4%) cases, and MDR-TB was found in 8/223 (3.6%) cases. We found significantly different amounts of DR-TB by location (18.18% in the south of the country versus 3.91% in the north central region [ P < 0.01]). Furthermore, RIF resistance was genetically distinct, suggesting possible location-specific strains are responsible for the transmission of drug resistance ( P < 0.04). Finally, GenoType MTBDRplus correctly identified the drug-resistant samples compared to sequencing in 96.8% of cases. We found that total DR-TB in HIV-infection is high and that transmission of drug-resistant TB in HIV-infected patients in Nigeria is higher than predicted.

Several genotypes of Mycobacterium with varied degree of virulence and drug resistance exist globally. Grouping of the genotypes into principal genetic groups is critical for tuberculosis (TB) control. This study determined the suitability of polymerase chain reaction- restriction enzyme analysis (PCR-REA) technique as an epidemiological typing tool and a technique for drug resistance surveillance of TB. Seventy-five Mycobacterium tuberculosis isolates from National TB Reference Laboratory, Nigeria Institute of Medical Research (NIMR) Yaba, Lagos, Nigeria were identified and their susceptibility to isoniazid and rifampicin determined using Genotype MTBDRplus technique. Isolates were further confirmed by polymerase chain reaction (PCR) with primer specific to 65 kDa heat shock protein (hsp65) of Mycobacterium species, Fingerprinting was done by restriction enzyme (Bst Ell) analysis. Of the 75 isolates studied, 5(6.7%) were multi-drug resistant (MDR)- TB, 11(14.7%) and 7(9.3%) were mono-resistant to rifampicin and isoniazid respectively. PCR-REA profile revealed 3 different patterns. Majority (96.2%) of the susceptible strains and 100% each of mono-resistant and MDR -TB yielded 2 fragments of 250 and 120 bp. Our findings demonstrated that hsp65 kDa based molecular genotyping technique is a rapid, simple and easy-to-interpret method and may be used as a predictor of M. tuberculosis genetic variation. Key words: Genotyping, Mycobacterium tuberculosis, heat shock protein, restriction enzyme analysis, drug susceptibility.

Issues: Quality-management systems (QMS) are uncommon in clinical laboratories in Nigeria, and until recently, none of the nation’s 5 349 clinical laboratories have been able to attain the certifications necessary to begin the process of attaining international accreditation. Nigeria’s Human Virology Laboratory (HVL), however, began implementation of a QMS in 2006, and in 2008 it was determined that the laboratory conformed to the requirements of ISO 9001:2000 (now 2008), making it the first diagnostic laboratory to be certified in Nigeria. The HVL has now applied for the World Health Organization (WHO) accreditation preparedness scheme. The experience of the QMS implementation process and the lessons learned therein are shared here.Description: In 2005, two personnel from the HVL spent time studying quality systems in a certified clinical laboratory in Dakar, Senegal. Following this peer-to-peer technical assistance, several training sessions were undertaken by HVL staff, a baseline assessment was conducted, and processes were established. The HVL has monitored its quality indicators and conducted internal and external audits; these analyses (from 2007 to 2009) are presented herein.Lessons learned: Although there was improvement in the pre-analytical and analytical indicators analysed and although data-entry errors decreased in the post-analytical process, the delay in returning laboratory test results increased significantly. There were several factors identified as causes for this delay and all of these have now been addressed except for an identified need for automation of some high-volume assays (currently being negotiated). Internal and external audits showed a trend of increasing non-conformities which could be the result of personnel simply becoming lax over time. Application for laboratory accreditation, however, could provide the renewed vigour needed to correct these non-conformities.Recommendation: This experience shows that sustainability of the QMS at present is a cause for concern. However, the tiered system of accreditation being developed by WHO–Afro may act as a driving force to preserve the spirit of continual improvement.

Background Orofacial clefts are the most common malformations of the head and neck, with a worldwide prevalence of 1 in 700 births. They are commonly divided into CL(P) and CP based on anatomic, genetic, and embryologic findings. A Nigerian craniofacial anomalies study (NigeriaCRAN) was set up in 2006 to investigate the role of gene-environment interaction in the origin of orofacial clefts in Nigeria. Subjects and Methods DNA isolated from saliva from Nigerian probands was used for genotype association studies and direct sequencing of cleft candidate genes: MSX1, IRF6, FOXE1, FGFR1, FGFR2, BMP4, MAFB, ABCA4, PAX7, and VAX1, and the chromosome 8q region. Results A missense mutation A34G in MSX1 was observed in nine cases and four HapMap controls. No other apparent causative variations were identified. Deviation from Hardy Weinberg equilibrium (HWE) was observed in these cases ( p = .00002). A significant difference was noted between the affected side for unilateral CL ( p = .03) and bilateral clefts and between clefts on either side ( p = .02). A significant gender difference was also observed for CP ( p = .008). Conclusions Replication of a mutation previously implicated in other populations suggests a role for the MSX1 A34G variant in the development of CL(P).

A total of 2,570 apparently healthy human immunodeficiency virus-negative adults from the six geopolitical zones in the country were enrolled in our study in 2006. The samples were assayed using the Cyflow technique. Data were analyzed using the Statistical Package for Social Scientists (SPSS). The majority (64%) of the participants had CD4 counts within the range of 501 to 1,000 cells/μl. The reference range for CD4 was 365 to 1,571 cells/μl, while the reference range for CD8 was 145 to 884 cells/μl.

Three hundred and eighty-one patients with High Risk Bronchopulmonary Symptoms [HRBS] who presented at Yaba and Lagos University Teaching Hospital [LUTH] chest clinics were screened for Acid Fast Bacilli [AFB] comparatively, using Zhiel Nielson [ZN] staining method with Direct Smear [DS], Concentrated Smear [CS] and Concentrated deposit culture isolates on Lowenstein Jensen [LJ] medium, between April, 2000 to May, 2002. The prevalence rates of 24.4%, 36.2% and 49.6% were recorded respectively. The culture method were shown to correlate well with both DS [94.7%] and CS [98.8%] for positive samples; thus, demonstrating high specificity of the methods. Contamination rate of 6% were recorded during the culture procedure. Eighty-one percent of all positive samples were from both “on-the-spot” and “early morning” samples, while early morning samples alone made an additional 16 [19%] positive samples. The age specific incidence rate of positive subjects yielded more bacilli count [47%] of +++ [i.e. more than 10 acid fast bacilli per field in at least 20 fields], when compared with the direct smears with only 6.5% of +++. Apart from demonstrating the superiority of concentrated smears over the direct smears for AFB rapid and cheap screening, we equally confirmed the usefulness of good, adequate and properly timed sputum samples collection for improved TB case detection in a resource poor setting. Digested sputum procedure is therefore, recommended for routine use in out TB diagnostic centers as minimal training is only required. NQJHM Vol. 14 (1) 2004: pp. 8-10

There is an understanding that greater availability of HIV treatment for the 40.3 million people currently infected with HIV is a humanitarian imperative that could prolong the lives of millions, restore economic productivity, and stabilise societies in some of the world's hardest-hit regions. The Nigerian government recognises that the country has the third highest burden of infection, with people living with HIV estimated to total 4.0 million, and so in 2002 commenced the implementation of one of Africa's largest antiretroviral (ARV) treatment programmes. A successful ARV programme requires that all components of a functional management system be put in place for effective and efficient functioning. This would include logistics, human resources, financial planning, and monitoring and evaluation systems, as well as sustainable institutional capacities. The Nigerian national ARV treatment training programme was conceived to meet the human resource needs in hospitals providing ARV therapy. This paper reports on the evaluation of the training programme. It examines knowledge and skills gained, and utilisation thereof. Recommendations are made for improved training effectiveness and for specific national policy on training, to meet the demand for scaling up therapy to the thousands who need ARV. Keywords: ARV, training, evaluation, HIV, health care providerJournal of Social Aspects of HIV/AIDS Research Initiatve Vol. 3 (3) 2006: pp. 488-502

Definitive diagnosis of HIV infection in infants18 months of age who were born to HIV-infected mothers is still posing some difficulty in Nigeria and other developing countries. Within this age definitive diagnosis can only be carried out by antigen based techniques which are indeed not available in these developing countries. This has resulted in the absence of authoritative data on the rate of mother-to-child transmission in these countries. Nigeria inclusive. The present pilot study was therefore carried out to generate some information on the rate of mother to child transmission in Nigeria using the PCR technique. Plasma samples were obtained from 68 children of both sexes less than 18 months of age and who were born to HIV infected mothers. The samples were collected from two pediatric departments. in Lagos and in Benin. The presence of HIV 1 RNA in each of the samples. was determined using the Amplicor Monitor V 1.5 technique (Roche Diagnostics). Data showed that HIV-1 RNA was detected in 15 of the 68 samples tested. This gave an HIV-1 RNA detection rate of 22%. Among women who had some intervention, the rate of transmission of infection was 11% while the rate among those without intervention was 30%. The 22% transmission rate recorded in this study is close to the range of 25 to 35% that has been reported in several developed and a few developing countries. A multicenter nationwide study will still be needed to determine the national mother to child transmission rate in Nigeria.

Objective : To compare the efficacy of the Capcellia assay and the Dynabeads technique against the FACScount technique in the estimation of CD4 T-lymphocytes within a Nigerian setting. Design : Prospective study. Setting : Urban area in Nigeria. Subjects: Ninety seven subjects (51 HIV seronegative and 46 HIV seropositive adults)seen in the blood banks and two out patient clinics in Lagos within the study period. Main Outcome Measures : Dynadeads technique with a higher correlation coefficient is a feasible alternative to the FACScount method. Results : There was an overall correlation coefficient of r = 0.75 for CD4 cell counts as determined by the Dynabeads technique in comparison with the FACScount method. Also, an overall correlation coefficient of r = 0.17 for CD4 cell counts as determined by the Capcellia technique against the FACScount method. Conclusion : Dynabeads technique is simple to carry out and cheaper in terms of demand for human expertise and infrastructural requirements than the FACScounts. Therefore, it was recommended for use in the laboratory for monitoring of ARV therapy in Nigeria and any other resource poor country. East African Medical Journal Vol. 83(4) 2006: 105-111

The objective was to evaluate treatment outcome in the first 12 months among HIV-positive patients managed with a combination of nevirapine + stavudine + lamivudine under the current national antiretroviral (ARV) program in Nigeria. This was a prospective observational cohort study on 50 ARV-naive patients who met the inclusion criteria for the program and had given informed consent. All patients were in stage 2 or stage 3 periods of infection based on World Health Organization clinical classification. The patients were treated with the generic brands of ARVs and treatment consisted of oral nevirapine (Nevimal Cipla Mumbai India) 200 mg daily lamivudine (Lamivir Cipla) 150 mg twice daily and stavudine (Stavir Cipla) 40 mg twice daily. Prior to initiation of treatment the clinical history and baseline data for each patient were documented. The levels of plasma HIV-1 RNA CD4+ cell counts frequency of opportunistic infections and estimated body mass index were recorded at baseline and subsequently at intervals during treatment. Data obtained at the various sampling times for each parameter were compared against their baseline values. Data on the plasma HIV-1 RNA levels indicated that between baseline and week 24 the median viral load of the patients decreased by 1.79 log10 copies/mL. Equally between baseline and week 48 the median CD4+ cell counts increased by 186 3 106 cells/L the frequency of opportunistic infections decreased by 82% the median body mass index increased by 4.8 kg/m2 and 36% experienced side effects which were minor and transient. The most prevalent side effect recorded was skin rash associated with nevirapine. Good adherence to this triple regimen was recorded in .85% of the patients. The overall results within the 12-month treatment period indicated an effective suppression of viral replication the reconstitution of the immune system and improvement of the physical well-being of the study population. Though there may be differences in global distribution of the infecting HIV-1 subtypes the clinical and biologic results of this study compared favorably to those documented in cohorts treated with branded and generic ARV drugs in some developed and developing countries. The cumulative data in this study further confirmed that the correct use of generic brands of ARVs is a feasible option in HIV care and support programs in resource-poor countries. (authors)

Immune dysregulation in terms of hypergammaglobulinaemia and hypogammaglobulinaemia was part of definition of immunodeficiency that characterized AIDS. This results in either raised level of total plasma/serum protein or low level of plasma/serum protein depending on which component of immune dysregulation predominates. This study examined the level of total serum proteins and globulins in HIV infected Nigerians. 64 patients with HIV infection and 10 apparently healthy subjects were recruited from 3 hospitals in Lagos Metropolis. They were examined for the presence of TB and malaria. Serum total protein, albumin and creatinine levels were determined using commercially available kits while the globulin level was computed. Level of immunodeficiency was also determined by CD4+ cell count, using the FACS count method. Data obtained from the patients were compared with values obtained from 10 healthy controls; using appropriate statistical tests on EPI-INFO V 6.0. Results showed that hypergammaglobulinaemia is the major immune dysregulation found with globulin level of 7.4g/d1 as compared with 2.9g/d1 in healthy control though it was not statistically significant. Total serum protein was also higher in HIV infected patients than the controls. The serum globulin was found to correlate negatively with CD4 cell count (r = -0.28, P = 0.0067). Serum creatinine appears not to differ significantly between control and infected patients although the creatinine levels has a positive correlation to plasma globulin levels (r = 0.45, p = 0.001). In conclusion hypergammaglobulinaemia was found to be common in HIV infected Nigerians. This was associated with raised level of plasma creatinine. KEY WORDS: Serum proteins, Creatinine, Albumin, Globulin and HIV Nigerian Journal of Health and Biomedical Sciences Vol.3(2) 2004: 69-72

The objective of this study was to compare the sensitivity and specificity of smear and culture methods with rapid serological EIA myco kits manufactured by Omega diagnostics, for the early detection of Mycobacterium tuberculosis (MTB) complex. Sera from various categories of smear and culture results were compared with the result of 38KDa, 16KDa and purified protein for IgA, IgM and IgG antibodies with sensitivity of 4%, 24% and 76%, respectively and with specificity of100% for IgG in Smear and Culture Positive (S + + - + - - + + + + + + + Key Words: Tuberculosis, diagnosis, comparative, specificity, sensitivity, culture and serological technique. Afr. J. Clin. Exper. Microbiol. 2004; 5(2): 182 – 188.

The study examined a possible association between HIV infection and conventional sexually transmitted diseases (STDS) in a population of 700 patients seen in some hospitals and clinics in Lagos State between November 1997 and December 1999. The patients were drawn mainly from LUTH and Jolad hospitals in Lagos State. In these hospitals, patients who presented with symptoms of STDS were screened clinically and microbiologically for agents of STDS and HIV antibodies. Screening was carried out using conventional methods. A total of 150 (21.5%) were found positive for various STDS while 550 (78.5%) were negative Also, 109 (15.8%) were sero-positive for HIV while 591 (84.4%) were sero-negative. The frequency of STDS diagnosed were, Treponema pallidum, 38(25.3%), Neisseria gonorrhoea 3(2.0%), Chlamydia trachomatis 26(17.3), Hepatitis B virus 60(40.0%) Staphylococcus aureaus, 20 (13.3%) and Candida albicans 3(2.0%). Data showed that Syphillis was the most prevalent STDS diagnosed while Calbicans and N. gonorrhoea are the least. Amongst the 150 (21.5%) patients positive with STDS, 82(54.65%) were found to be positive for HIV antibodies. The remaining 68(45.3%) patients were negative for HIV. The difference in sero-prevalence on the true group of patients rates was significant. The higher rate in the STDS patients strongly suggest some association between HIV infections andSTDS amongst the patients studied p = 0.05. It was also recorded that HIV-1 infection is four times more prevalent than HIV-2 in these patients.

Severe falciparum malaria, with its associated hyperpyrexia, distorts plasma levels of large neutral amino acids (NAA) and consequently, brain uptake of individual NAA. Since brain levels of NAA determine cerebral synthesis of monoamines (serotonin, histamine, catecholamines), we measured plasma concentrations of NAA, and also plasma histamine (Hm) in children with falciparum malaria and in uninfected controls. Malaria elicited a marked (P < 0.025) increase in plasma histidine (His) with a 5-fold significant (P < 0.001) elevation in histamine, as well as a 2.5-fold increase (P < 0.005) in plasma phenylalanine (Phe), with no changes in the other NAA. Using kinetic parameters of NAA transport at human blood-brain barrier (BBB), we showed that malaria significantly altered calculated brain uptake of His (+30%), Phe (+96%), Trp (-30%) and Ile (-27%), with no change in the other NAA, compared with controls. Our data suggested enhanced cerebral synthesis of Hm with impaired production of serotonin and the catecholamines in the patients, and therefore, the need to evaluate the encephalopathy in severe malaria within the context of abnormalities in metabolism of Hm and other monoamines resulting from imbalance in plasma levels of the large neutral amino acids. Of clinical relevance also is the impaired inactivation of increased brain Hm by antimalarials such as the widely used aminoisoquinolines leading to elevated brain levels of imidazole-4-acetic acid (IAA), a potent inducer of a sleep-like state often accompanied by seizures, analgesia, decreased blood pressure and other effects.

Brain monoamine levels may underlie aspects of the cerebral component of falciparum malaria. Since circulating amino acids are the precursors for brain monoamine synthesis, we measured them in malaria patients and controls. Malaria elicited significantly elevated plasma levels of phenylalanine, particularly in comatose patients, with the Tyr/Phe (%) ratio reduced from 83.3 in controls to 39.5 in infected children, suggesting an impaired phenylalanine hydroxylase enzyme system in malaria infection. Malaria significantly increased the apparent K(m) for Trp, Tyr and His, with no effect on K(m)(app) for Phe. Using the kinetic parameters of NAA transport at the human blood-brain barrier, malaria significantly altered brain uptake of Phe (+96%), Trp (-28%) and His (+31%), with no effect on Tyr (-8%), compared with control findings. Our data suggest impaired cerebral synthesis of serotonin, dopamine and norepinephrine, and enhanced production of histamine, in children with severe falciparum malaria.

Noma (cancrum oris) is an infectious disease which destroys the oro-facial tissues and other neighboring structures in its fulminating course. It affects predominantly children aged 2-16 years in sub-Saharan Africa where the estimated frequency in some communities may vary from one to seven cases per 1000 children. The key risk factors are poverty, malnutrition, poor oral hygiene, deplorable environmental sanitation, close residential proximity to livestock, and infectious diseases, particularly measles. Malnutrition acts synergistically with endemic infections in promoting an immunodeficient state, and noma results from the interaction of general and local factors with a weakened immune system as the common denominator. Acute necrotizing gingivitis (ANG) is considered the antecedent lesion. Current studies suggest that evolution of ANG to noma requires infection by a consortium of microorganisms with Fusobacterium necrophorum and Prevotella intermedia as the suspected key players. Without appropriate treatment, mortality rate is 70-90%. Survivors suffer the two-fold affliction of oro-facial disfigurement and functional impairment. Reconstructive surgery of the resulting deformity is time-consuming and financially prohibitive for the victims who are poor.

The microbiologic history of noma was reviewed. Studies have associated the disease process with large numbers of fusiform bacilli and spirochetal organisms. In order to study the microbiology of the staging and infection periods of noma 62 Nigerian children, aged 3-14 years, 22 children had acute necrotizing ulcerative gingivitis (ANUG) and were also malnourished, 20 exhibited no acute necrotizing ulcerative gingivitis but were malnourished and 20 were free of acute necrotizing ulcerative gingivitis and in good nutritional state) were evaluated for the presence of viruses and oral microorganisms. The ANUG cases in the malnourished children had a higher incidence of Herpesviridae, the main virus being detected was cytomegalovirus. There were more anaerobic microorganisms recovered, with Prevotella intermedia as the predominant isolate, in the malnourished children as compared to the healthy children. A study of the predominant microflora in active sites of noma lesions was carried out in eight noma patients, 3-15 years of age, in Sokoto State, northwestern Nigeria. Fusobacterium necrophorum was recovered from 87.5% of the noma lesions. Oral microorganisms isolated included Prevotella intermedia, alpha-hemolytic streptococci and Actinomyces spp. which were isolated from 75.0, 50.0 and 37.5% of the patients, respectively. Peptostreptococcus micros, Veillonella parvula, Staphylococcus aureus and Pseudomonas spp. were each recovered from one lesion. All strains were observed to be sensitive to all of the antibiotics tested with the exception of one strain of P. intermedia which showed resistance to penicillin. The pathogenic mechanisms of F. necrophorum as a trigger organism were discussed. The isolation from human noma lesions of F. necrophorum, a pathogen primarily associated with animal diseases, may have important etiologic and animal transmission implications.

This study showed that impoverished Nigerian children at risk for cancrum oris (noma) had significantly reduced plasma concentrations of zinc (< 10.8 micromol/L), retinol (< 1.05 micromol/L), ascorbate (< 11 micromol/L), and the essential amino acids, with prominently increased plasma and saliva levels of free cortisol, compared with their healthy counterparts. The nutrient deficiencies, in concert with previously reported widespread viral infections (measles, herpesviruses) in the children, would impair oral mucosal immunity. We postulate, subject to additional studies, that evolution of the oral mucosal ulcers including acute necrotizing gingivitis to noma is triggered by a consortium of microorganisms of which Fusobacterium necrophorum is a key component. Fusobacterium necrophorum elaborates several dermonecrotic toxic metabolites and is acquired by the impoverished children via fecal contamination resulting from shared residential facilities with animals and very poor environmental sanitation.

To establish the prevalence of HIV antibodies in patients with pulmonary tuberculosis, 536 new cases presenting with symptoms of bronchopulmonary disorders were randomly selected from the six referral chest clinics in Lagos and screened for tuberculosis and HIV infections. Sputum and serum samples were obtained from all the patients. The sputum samples were examined for acid-fast bacilli (AFB) by both microscopy and culture. The sera were screened for HIV-1 and HIV-2 antibodies by ELISA and confirmed by Western blot (WB). Of the 536 cases studied, 188 (35%) were positive for AFB while 13 (2.4%) were seropositive for HIV. Correlation between the AFB and HIV results revealed that 10 (5.3%) of the 188 AFB positives were also seropositive for HIV as compared to 3 (0.9%) in the 348 AFB negative cases. The difference in the HIV seroprevalence rates in the two groups was statistically significant (P0.001). The recorded higher frequency of HIV infections in the AFB positives strongly suggested some level of interaction between TB and HIV infections in Lagos. Infections with HIV-2 were more prevalent than HIV-1 in the patients with HIV and TB. No case of dual infection with HIV-1 and HIV-2 was recorded in this group of patients. However, in the 3 HIV-seropositive patients within the control group (non-tuberculosis patients), 2 (67%) were positive for both HIV-1 and HIV-2 while 1 (33%) was positive for HIV-2 only. Mycobacterium tuberculosis (70%), M. avium (20%) and M. kansasii (10%) were the mycobacteria strains isolated from the HIV/TB infected patients.(ABSTRACT TRUNCATED AT 250 WORDS)

The extent and pattern of drug resistance among previously treated tuberculosis patients was investigated. Ninety-six patients with a total treatment duration of between 6 and 18 months and still smear and culture positive were examined. Treatment was either continuous or in intermittent blocks. Drug susceptibility tests on strains of tubercle bacilli isolated from the patients were performed against isoniazid, streptomycin, p-aminosalicylic acid, ethambutol and rifampicin by the proportion method using LJ medium without potato starch. A total of 56% of the strains were resistant to one or more of the drugs tested. Resistance to isoniazid (38%) and streptomycin (29%) was most common. A significant finding in the study was the low level of resistance to rifampicin (2%) and ethambutol (3%). A relationship between the incidence of drug resistance and the nature and duration of previous treatment appeared likely since susceptible strains were isolated more often from patients with continuous treatment than from patients on intermittent blocks of long-course regimens. It is therefore suggested that the introduction of better supervision of drug taking and the adoption of continuous short-course regimens on a nationwide level will contribute immensely towards the reduction of the drug resistance problems in Nigeria as well as in other developing countries.

A total of 2784 cases with persistent symptoms of lower respiratory tract infections were seen in four of the chest clinics in Lagos between January and June of 1983. Of these, 668 were randomly selected and screened for pulmonary tuberculosis. Repeated sputum samples from the 668 patients were cultured on Lowenstein-Jenseen slopes and 102 pure mycobacterial isolates were obtained. The isolates were differentiated into tubercle bacilli, bovine or atypical mycobacteria on the basis of results obtained from nine identification tests. Of the 102 mycobacterial isolates obtained, 87 (85%) were identified as Mycobacterium tuberculosis, four (4%) as M. bovis and 11 (11%) as atypical mycobacteria. Among this atypical group, six were classified as M. avium, four as M. kansasii and one as M. fortuitum. The study showed the involvement of bovine and atypical mycobacteria in pulmonary infections in Lagos. Tuberculosis-like diseases produced by some of the atypical group are resistant to most of the conventionally used anti-tuberculosis drugs. The need for adequate bacteriological analysis in current-day diagnosis and treatment of pulmonary tuberculosis is therefore highlighted since the 11 atypical cases seen in this study would otherwise have been diagnosed and treated as cases of classical tuberculosis.

The pattern of loss of histamine-sensitizing factor (HSF) during antigenic modulation of Bordetella pertussis in Hornibrook medium was examined. The aim was to determine the possible underlying mechanism involved in modulation. Normal (X-mode) B. pertussis cells were grown in Hornibrook medium in which 0.5% (w/v) NaCl had been replaced with 0.5% (w/v) MgSO4. 7H2O (C-medium). At various time intervals during growth, the viable cell numbers and optical densities of both cultures in the X- and C-media were estimated. Whole cells were harvested from the cultures at the same time intervals and aliquots from the cultures were assayed for the levels of their histamine-sensitizing properties. Correlation of the increase in viable cell numbers with rate of loss of histamine-sensitizing activity in both the cells and whole cultures indicated that components responsible for the histamine-sensitizing activity were not synthesized during modulation. Moreover, the loss of HSF from B. pertussis cells was faster than can be explained by dilution of the original factor in the inoculum among progeny cells. Modulation may involve cessation of synthesis and selective degradation or denaturation of some envelope polypeptides immediately upon inoculation of normal X-mode B. pertussis cells into C-medium.

SUMMARY. Modulation of Bordetella pertussis was induced by growth in Hornibrook medium with a high content of magnesium sulphate (C-medium); four pathophysiological activities in the cells or in the whole culture were measured at intervals. Modulation, shown by the extensive loss of protective antigen, histamine-sensitising factor, leuko-cytosis-promoting factor, heat-labile toxin and X-mode specific envelope proteins, occurred during the first 10 h of incubation of X-mode cells in C-medium at 35°C and before the onset of the logarithmic phase of growth. The rapidity of these losses was greater than could be explained by a simple growth-dilution effect and did not appear to be due to release of activity into the culture fluid. It seemed, therefore, that there was selective destruction of pathophysiological activities as well as cessation of synthesis. The activities appeared to be lost at different rates. Mouse-protective activity was lost more rapidly than histamine-sensitising and leukocytosis-promotmg activities; heat-labile toxicity was lost more slowly or less completely. Modulation was shown to be easily reversed.

Background : The Nigerian Institute of Medical Research houses two reference laboratories: the virology and tuberculosis laboratories. Both were enrolled in the Strengthening Laboratory Management Toward Accreditation (SLMTA) programme. Objective: To describe the impact of SLMTA and discuss factors affecting the results, with an emphasis on mentorship. Methods : The SLMTA programme was implemented from April 2010 through November 2012. Participants attended three workshops and executed quality improvement projects; laboratory auditors evaluated performance using a standard checklist. The virology laboratory did not receive mentorship; however, the tuberculosis laboratory had an international mentor who visited the laboratory four times during the programme, spending two to four weeks embedded within the laboratory during each visit. Results : There was an overall improvement in the performance of both laboratories, with the virology laboratory increasing 13% (from 80% at baseline to 93% at exit audit) and the tuberculosis laboratory increasing 29% (from 66% to 95%). These scores were maintained nine months later at the surveillance audit. Conclusion : The SLMTA programme resulted in improved and sustained quality management performance for both laboratories. Mentoring was a possible factor in the substantial improvement made by the tuberculosis laboratory and should be considered in order to augment the training received from the SLMTA workshops.

In this study, 65 patients are screened for Salmonella typhi by conventional culture and the Widal test. In addition, the patients undergo full blood count are screened for malaria parasites. Of the 65 patients, 50 report febrile conditions, while the remaining 15 are used as a control population. In the febrile group, 13 (26%) were positive for S. typhi, while in the control group only one (7%) was positive for S. typhi. Overall, 36 (64.3%) patients had malaria parasites. Patients with a higher O antibody titre (> or = 1 in 80) by Widal test were found to have consumed both tap water and pure water. More females (10/14; mean age: 33) had typhoid fever as a result of S. typhi infection, the majority of which were isolated from stool samples (57%). Nine of the isolates were also positive for malaria parasites, seven of which were in the trophozoite stage. Plasmodium falciparum was the predominant parasite (78%), the remainder being P. malariae. The majority of patients (12/14) with typhoid fever had normal PCV values. In conclusion, it is recommended that tests for the diagnosis of typhoid fever in Nigeria should include malaria parasites, S. typhi culture from faecal samples, and the Widal test.