Nevirapine plasma concentration is associated with virologic failure and the emergence of drug-resistant mutations among HIV patients in Kenya: A cross sectional study.

This study aimed to determine the association between the plasma concentration of nevirapine (NVP) and clinical outcomes. In this cross-sectional study, sociodemographic and clinical data were collected from 233 HIV patients receiving NVP-based first-line antiretroviral therapy (ART) regimens in Nairobi, Kenya. The mean age was 41.2 (SD ±â€…11.9) years. Fifty-four (23.2%) patients had virological failure (>1000 copies/mL), whereas 23 (9.9%) were infected with drug-resistant HIV strains. Eleven patients had nucleoside reverse transcriptase inhibitor resistance mutations, including M184V and T215Y, whereas 22 had non-nucleoside reverse transcriptase inhibitor resistance mutations, including G190A, K103N, V106A, Y181C, A98G, and Y188L. The median NVP plasma concentration was 6180 ng/mL (IQR 4444-8843 ng/mL), with 38 (16.3%) patients having suboptimal NVP plasma levels of <3400 ng/mL. The majority 23 of the 38 (60.5%) patients with NVP Cmin < 3400 ng/mL were significantly infected with drug-resistant HIV virus (P = .001). In the multivariate analysis, the time taken to arrive at the ART clinic (ß -11.1, 95% CI -21.2 to -1.1; P = .031), higher HIV viral load (ß -2008, 95% CI -3370.7 to -645.3; P = .004), and the presence of HIV drug resistance mutation (ß 3559, 95% CI 2580.8-4537.2; P = .0001) were associated with NVP plasma concentration. A significant proportion of patients receiving the NVP-based regimen had supra- and sub-therapeutic plasma concentrations. Higher HIV viral load and the presence of HIV drug-resistant mutations are important factors associated with NVP plasma concentrations.

Effects of cytochrome P450 2B6 and constitutive androstane receptor genetic variation on Efavirenz plasma concentrations among HIV patients in Kenya.

The effects of genetic variation of cytochrome P450 2B6 (CYP2B6) and constitutive androstane receptor (CAR) on efavirenz (EFV) plasma concentration was evaluated among 312 HIV patients in Nairobi Kenya. The EFV plasma concentration at steady-state were determined using ultra-high-performance liquid chromatography with a tandem quadruple mass spectrometer (LC-MS/MS). Thirteen CYP2B6 (329G>T, 341T>C, 444 G>T/C, 15582C>T, 516G>T, 548T>G, 637T>C, 785A>G, 18492C>T, 835G>C, 1459C>T and 21563C>T) and one CAR (540C>T) single nucleotide polymorphisms (SNPs) were genotyped using real-time polymerase chain reaction. HIV drug resistance mutations were detected using an in-house genotypic assay. The EFV concentration of patients ranged from 4 ng/mL to 332697 ng/mL (median 2739.5 ng/mL, IQR 1878-4891.5 ng/mL). Overall, 22% patients had EFV concentrations beyond therapeutic range of 1000-4000 ng/mL (4.5%% < 1000 ng/mL and 31.7% > 4000 ng/mL). Five SNPs (15582C>T, 516G>T, 785A>G, 983T>C and 21563C>T) were associated with higher EFV plasma concentration while 18492C>T with lower EFV plasma concentration (p<0.05). Strong linkage disequilibrium (LD) was observed for 15582C>T, 516G>T, 785A>G, 18492C>T, 983T>C, 21563C>T, 1459C>T and CAR 540C>T. Sixteen haplotypes were observed and CTGCTTCC, CTGCTTCT, TTGCTTCT and CGACCCCT were associated with high EFV plasma concentration. In multivariate analysis, factors significantly associated with EFV plasma concentration included; the presence of skin rash (ß = 1379, 95% confidence interval (CI) = 3216.9-3416.3; p < 0.039), T allele of CYP2B6 516G>T (ß = 1868.9, 95% CI 3216.9-3416.3; p < 0.018), the C allele of CYP2B6 983T>C (ß = 2638.3, 95% CI = 1348-3929; p < 0.0001), T allele of CYP2B6 21563C>T (ß = 1737, 95% CI = 972.2-2681.9; p < 0.0001) and the presence of 5 to 7 numbers of SNPs per patient (ß = 570, 95% CI = 362-778; p < 0.0001) and HIV viral load ≤1000 cells/mL (ß = -4199.3, 95% CI = -7914.9 --483.6; p = 0.027). About 36.2% of the patients had EFV plasma concentrations beyond therapeutic window, posing high risk of treatment failure or toxicity. The SNPs of CYP2B6 516G>T, CYP2B6 983T>C, 21563C>T, presence of higher numbers of SNPs per patient and haplotypes CTGCTTCC, CTGCTTCT, TTGCTTCT and CGACCCCT could efficiently serves as genetic markers for EFV plasma concentration and could guide personalization of EFV based ART treatment in Kenya.

Association between social psychological status and efavirenz and nevirapine plasma concentration among HIV patients in Kenya.

HIV-related stigma, lack of disclosure and social support are still hindrances to HIV testing, care, and prevention. We assessed the association of these social-psychological statuses with nevirapine (NVP) and efavirenz (EFV) plasma concentrations among HIV patients in Kenya. Blood samples were obtained from 254 and 312 consenting HIV patients on NVP- and EFV-based first-line antiretroviral therapy (ART), respectively, and a detailed structured questionnaire was administered. The ARV plasma concentration was measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). There were 68.1% and 65.4% of the patients on NVP and EFV, respectively, who did not feel guilty for being HIV positive. The disclosure rates were approximately 96.1% and 94.6% of patients on NVP and EFV, respectively. Approximately 85% and 78.2% of patients on NVP and EFV, respectively, received social support as much as needed. There were 54.3% and 14.2% compared to 31.7% and 4.5% patients on NVP and EFV, respectively, with supratherapeutic and suboptimal plasma concentrations. Multivariate quantile regression analysis showed that feeling guilty for being HIV positive was associated with increased 954 ng/mL NVP plasma concentrations (95% CI 192.7 to 2156.6; p = 0.014) but not associated with EFV plasma concentrations (adjusted ß = 347.7, 95% CI = - 153.4 to 848.7; p = 0.173). Feeling worthless for being HIV positive was associated with increased NVP plasma concentrations (adjusted ß = 852, 95% CI = 64.3 to 1639.7; p = 0.034) and not with EFV plasma concentrations (adjusted ß = - 143.3, 95% CI = - 759.2 to 472.5; p = 0.647). Being certain of telling the primary sexual partner about HIV-positive status was associated with increased EFV plasma concentrations (adjusted ß 363, 95% CI, 97.9 to 628.1; p = 0.007) but not with NVP plasma concentrations (adjusted ß = 341.5, 95% CI = - 1357 to 2040; p = 0.692). Disclosing HIV status to neighbors was associated with increased NVP plasma concentrations (adjusted ß = 1731, 95% CI = 376 to 3086; p = 0.012) but not with EFV plasma concentrations (adjusted ß = - 251, 95% CI = - 1714.1 to 1212.1; p = 0.736). Obtaining transportation to the hospital whenever needed was associated with a reduction in NVP plasma concentrations (adjusted ß = - 1143.3, 95% CI = - 1914.3 to - 372.4; p = 0.004) but not with EFV plasma concentrations (adjusted ß = - 6.6, 95% CI = - 377.8 to 364.7; p = 0.972). HIV stigma, lack disclosure and inadequate social support are still experienced by HIV-infected patients in Kenya. A significant proportion of patients receiving the NVP-based regimen had supra- and subtherapeutic plasma concentrations compared to EFV. Social-psychological factors negatively impact adherence and are associated with increased NVP plasma concentration compared to EFV.

Microbiological Safety of Fresh Tilapia ( Oreochromis niloticus) from Kenyan Fresh Water Fish Value Chains.

This study evaluated the microbiological safety of fresh Nile tilapia ( Oreochromis niloticus) from Kenyan fresh water fish value chains. One hundred seventy-six fish samples were analyzed. The microbial counts of hygiene indicators, total viable aerobic count (TVC), total coliforms, and fecal coliforms isolated by using culture techniques were enumerated, and microbial pathogens present in the fish samples were identified and characterized by using molecular methods. The diversity of bacterial isolates was determined by using the Shannon-Weaver diversity index. The mean of TVC in the samples was 4.44 log CFU/g. A comparison with the European Commission and International Commission on Microbiological Specifications for Foods standards showed two fish samples had counts above the 5.00 log CFU/g limit for TVC, and all the fish samples had total coliform and fecal coliform counts above 2.00 and 1.00 log CFU/g, respectively. Pathogenic strains, including Shiga toxin-producing and enteropathogenic Escherichia coli, Listeria monocytogenes, Yersinia enterocolitica, Klebsiella pneumoniae, and Salmonella enterica, were identified in the fish samples. The diversity of 1,608 bacterial isolates was higher in semiregulated chains than unregulated chains. The diversity was also high at the retail stage of the fish value chain. In conclusion, fresh Nile tilapia samples were above some of the set food safety standards and may be a source of foodborne pathogens. Further microbial risk assessment for detected pathogens is recommended to further support public health protection, taking into account growth, inactivation through cooking, processing, survival, and consumption.

Etiology of bacterial meningitis: a cross-sectional study among patients admitted in a semi-urban hospital in Nairobi, Kenya.

INTRODUCTION: bacterial meningitis, responsible for childhood morbidity and mortality, can also lead to permanent neurological disability among survivors. This study conducted from January to December, 2015 used standard bacteriological and molecular methods to investigate the etiology of three common causes of bacterial meningitis among hospitalized patients admitted at a semi-urban hospital in Nairobi, Kenya. METHODS: a total of 196 patients admitted at Mama Lucy Kibaki with clinically diagnosed meningitis were recruited into this cross-sectional study. Participants' information was collected through patient interviews and abstraction of health records. Bacterial culture, gram stains and multiplex polymerase chain reaction (PCR) were used to investigate causes of bacterial meningitis from cerebrospinal fluid (CSF) samples. Characteristics such as age, gender, occupation, underlying conditions of patients with laboratory confirmed bacterial meningitis infection are described. RESULTS: among the 196 patients diagnosed with bacterial meningitis, the median age was 1 year (range 1 to 36 years) with 87.2% aged 1 to 4 years; 54.6% were males. Using PCR, 22 out of 196 (11.2%) samples had evidence suggesting a bacterial infection. These included 12/22 (54.5%) S. pneumonia, 7/22 (31.8%) N. meningitides and 3/22 (13.6%) H. influenza. From bacterial culture, four of 196 (2.1%) samples grew S. pneumonia. All three samples found positive for H. influenza were from male patients aged between 1 to 4 years. CONCLUSION: of the three common causes evaluated, S. pneumonia was the most common cause of bacterial meningitis among patients from this region, particularly among infants. One older patient was diabetic, thereby highlighting the importance of pre-existing conditions. Although serotyping of bacteria was not done, under-vaccination might have played a role in the cases identified and ensuring complete and timely vaccination may prevent further cases of bacterial meningitis.

Fecal contamination of drinking water in Kericho District, Western Kenya: role of source and household water handling and hygiene practices.

Inadequate protection of water sources, and poor household hygienic and handling practices have exacerbated fecal water contamination in Kenya. This study evaluated the rate and correlates of thermotolerant coliform (TTC) household water contamination in Kericho District, Western Kenya. Culture and multiplex polymerase chain reaction (PCR) techniques were used to characterize TTCs. The disk diffusion method was used for antibiotic susceptibility profiling of pathogenic Escherichia coli. Out of the 103 households surveyed, 48 (46.6%) had TTC contaminated drinking water (TTC levels of >10 cfu/100 mL). Five of these households were contaminated with pathogenic E. coli, including 40% enteroaggregative E. coli, 40% enterotoxigenic E. coli, and 20% enteropathogenic E. coli. All these pathogenic E. coli strains were multidrug resistant to sulfamethoxazole/trimethoprim, ampicillin, tetracycline and ampicillin/sulbactam. Rural household locality, drinking water hand contact, water storage container cleaning practice, hand washing before water withdrawal, water source total coliforms <10 cfu/100 mL, temperature, and free chlorine levels were associated with TTC contamination of household drinking water. Significant proportions of household drinking water in Kericho District are contaminated with TTCs including with pathogenic multidrug-resistant E. coli. Source and household hygiene and practices contribute significantly to drinking water contamination.

HIV type 1 drug resistance patterns among patients failing first and second line antiretroviral therapy in Nairobi, Kenya.

BACKGROUND: The ever-expanding rollout of antiretroviral therapy in poor resource settings without routine virological monitoring has been accompanied with development of drug resistance that has resulted in limited treatment success. METHODS: A cross-sectional study with one time viral load was conducted during the period between 2012 and 2013 to determine treatment failure and drug resistance mutations among adults receiving first-line (44) (3TC_d4T/AZT_NVP/EFV) and second-line (20) (3TC/AZT/LPV/r) in Nairobi, Kenya. HIV-1 pol-RT genotyping for drug resistance was performed using an in-house protocol. RESULTS: A total of 64 patients were recruited (mean age 36.9 yrs.) during the period between 2012 and 2013 of the 44 adult patients failing first-line 24 (40.9%) had drug resistance mutations. Eight (8) patients had NRTI resistance mutations with NAMS M184V (54.2%) and K65R (8.4%) mutations being the highest followed by TAMs T215Y and K70R (12.5%). In addition, among patients failing second-line (20), six patients (30%) had NNRTI resistance; two patients on K103N and G190A mutations while V106A, Y184V, A98G, Y181C mutations per patient were also detected. However, for NRTI two patients had TAM T215Y. M184V mutation occurred in one patient. CONCLUSIONS: The study findings showed that HIV-1 drug resistance was significantly high in the study population. The detected accumulated resistance strains show that emergence of HIV drug resistance will continue to be a big challenge and should be given more attention as the scale up of treatment in the country continues.

Examination of a second region of the HIV type 1 genome reveals additional cases of superinfection.

HIV-1 superinfection may occur at a rate similar to that of initial infection, raising concerns for HIV-1 vaccine strategies predicated on eliciting immune responses similar to those in natural infection. Because of the high rate of recombination during HIV-1 replication, studies examining only one region of the HIV-1 genome are likely to miss cases of HIV-1 superinfection. We examined HIV-1 gag sequences from 14 high-risk Kenyan women in whom superinfection was not detected in a previous study of env sequences. We detected two additional cases of HIV-1 superinfection: one intersubtype superinfection that occurred between 1046 and 1487 days postinfection (DPI) and one intrasubtype superinfection that occurred between 341 and 440 DPI. Our results suggest that studies that examine only small genome regions may lead to underestimates of the risk of superinfection, highlighting the need for more extensive studies examining multiple regions of the HIV-1 genome.