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Featured researches published by Anne E. Asuquo.


Infection, Genetics and Evolution | 2012

Molecular epidemiology and genetic diversity of Mycobacterium tuberculosis complex in the Cross River State, Nigeria

Benjamin P. Thumamo; Anne E. Asuquo; Lydia N. Abia-Bassey; Lovett Lawson; Véronique Hill; Thierry Zozio; Nnamdi Emenyonu; Francis O. Eko; Nalin Rastogi

This study provides with a first insight on Mycobacterium tuberculosis complex epidemiology and genetic diversity in the Cross River State, Nigeria. Starting with 137 smear positive patients recruited over a period of 12months (June 2008 to May 2009), we obtained 97 pure mycobacterial isolates out of which 81 (83.5%) were identified as M. tuberculosis complex. Genotyping revealed a total of 27 spoligotypes patterns with 10 clusters (n=64% or 79% of clustered isolates, 2-32 isolates/cluster), with patients in the age group range 25-34 years being significantly associated with shared-type pattern SIT61 (p=0.019). Comparison with SITVIT2 database showed that with the exception of a single cluster (SIT727/H1), all other clusters observed were representative of West Africa; the two main lineages involved were LAM10-CAM (n=42/81% or 51.8%) of M. tuberculosis and AFRI_2 sublineage of Mycobacterium africanum (n=27/81% or 33.3%). Subsequent 12-loci MIRU typing resulted in a total of 13 SIT/MIT clusters (n=52 isolates, 2-9 isolates per cluster), with a resulting recent n-1 transmission rate of 48.1%. Available drug-susceptibility testing (DST) results for 58/81 clinical isolates revealed 6/58% or 10.4% cases of multiple drug-resistance (MDR); 5/6 MDR cases were caused by strains belonging to LAM10-CAM lineage (a specific cluster SIT61/MIT266 in 4/6 cases, and an orphan spoligotype pattern in 1/6 case). Additionally, MIT266 was associated with streptomycin resistance (p=0.016). All the six MDRTB isolates were concomitantly resistance to streptomycin and ethambutol; however, 4/6 MDR strains with identical MIRU patterns were characterized by consecutive strain numbers hence the possibility of laboratory cross contamination could not be excluded in 3/4 serial cases. The present preliminary study underlines the usefulness of spoligotyping and 12-loci MIRU-VNTRs to establish a baseline of circulating genotypic lineages of M. tuberculosis complex in Nigeria.


Tuberculosis Research and Treatment | 2012

Acid-Fast Bacilli Other than Mycobacteria in Tuberculosis Patients Receiving Directly Observed Therapy Short Course in Cross River State, Nigeria

Benjamin Thumamo Pokam; Anne E. Asuquo

The information on the contribution of non tuberculous mycobacteria (NTM) to mycobacterial infections in Africa is scarce due to limited laboratory culture for its isolation and identification. One hundred and thirty-seven sputum smear positive patients were recruited into a study on the molecular epidemiology of Mycobacterium tuberculosis in Cross River State. Following sputum culture, 97 pure isolates were obtained and identified using Capilia TB-Neo and further confirmed by the GenoType Mycobacterium CM kit. Of the 97 isolates, 81 (83.5%) isolates were Capilia TB-Neo positive while 16 (16.5%) were Capilia TB-Neo negative. Further confirmation with the GenoType Mycobacterium CM kit revealed that 4 (25%) of the 16 isolates belonged to NTM and included M. fortuitum I, M. fortuitum II/M magaritense, M. abscessus, and M. avium ssp. The remaining 12 (75%) Capilia TB-Neo negative isolates were not members of the genus Mycobacterium despite their AFB appearance. Six (33.3%) of the Capilia TB-Neo negative were from HIV positive tuberculosis patients. All subjects in this study were placed on DOTS shortly after the AFB results were obtained. The implication of isolation of 16.5% nontuberculous isolates further emphasizes the need for culture of sputum specimen especially in HIV positive patients prior to administration of antituberculosis therapy.


Scientific Reports | 2016

Abattoir-based estimates of mycobacterial infections in Cameroon

N. F. Egbe; Adrian Muwonge; Lucy M. Ndip; Robert Kelly; Melissa Sander; Vincent N. Tanya; V. Ngu Ngwa; Ian Handel; A. Novak; R. Ngandalo; K. L. Morgan; Anne E. Asuquo; B. M. de C. Bronsvoort

Mycobacteria cause major diseases including human tuberculosis, bovine tuberculosis and Johne’s disease. In livestock, the dominant species is M. bovis causing bovine tuberculosis (bTB), a disease of global zoonotic importance. In this study, we estimated the prevalence of Mycobacteria in slaughter cattle in Cameroon. A total of 2,346 cattle were examined in a cross-sectional study at four abattoirs in Cameroon. Up to three lesions per animal were collected for further study and a retropharyngeal lymph node was collected from a random sample of non-lesioned animals. Samples were cultured on Lowenstein Jensen media and the BACTEC MGIT 960 system, and identified using the Hain® Genotype kits. A total of 207/2,346 cattle were identified with bTB-like lesions, representing 4.0% (45/1,129), 11.3% (106/935), 23.8% (38/160) and 14.8% (18/122) of the cattle in the Bamenda, Ngaoundere, Garoua and Maroua abattoirs respectively. The minimum estimated prevalence of M. bovis was 2.8% (1.9–3.9), 7.7% (6.1–9.6), 21.3% (15.2–28.4) and 13.1% (7.7–20.4) in the four abattoirs respectively. One M. tuberculosis and three M. bovis strains were recovered from non-lesioned animals. The high prevalence of M. bovis is of public health concern and limits the potential control options in this setting without a viable vaccine as an alternative.


Scientific Reports | 2017

Molecular epidemiology of Mycobacterium bovis in Cameroon

N. F. Egbe; Adrian Muwonge; Lucy M. Ndip; Robert Kelly; Melissa Sander; Vincent N. Tanya; V. Ngu Ngwa; Ian Handel; A. Novak; R. Ngandalo; K. L. Morgan; Anne E. Asuquo; B. M. de C. Bronsvoort

We describe the largest molecular epidemiological study of Bovine Tuberculosis (bTB) in a sub-Saharan African country with higher spatial resolution providing new insights into bTB. Four hundred and ninety-nine samples were collected for culture from 201 and 179 cattle with and without bTB-like lesions respectively out of 2,346 cattle slaughtered at Bamenda, Ngaoundere, Garoua and Maroua abattoirs between 2012–2013. Two hundred and fifty-five M. bovis were isolated, identified and genotyped using deletion analysis, Hain® Genotype MTBC, spoligotyping and MIRU-VNTR. African 1 was the dominant M. bovis clonal complex, with 97 unique genotypes including 19 novel spoligotypes representing the highest M. bovis genetic diversity observed in Africa to date. SB0944 and SB0953 dominated (63%) the observed spoligotypes. A third of animals with multiple lesions had multiple strain infections. Higher diversity but little evidence of recent transmission of M. bovis was more common in Adamawa compared to the North-West Region. The Adamawa was characterised by a high frequency of singletons possibly due to constant additions from an active livestock movement network compared to the North-West Region where a local expansion was more evident. The latter combined with population-based inferences suggest an unstable and stable bTB-endemic status in the North-West and Adamawa Regions respectively.


The International Journal of Mycobacteriology | 2013

Multidrug resistance and demography of newly diagnosed tuberculosis patients in Cross River State, Nigeria

Benjamin Thumamo Pokam; Anne E. Asuquo; Lydia N. Abia-Bassey; Mandor B. Idasa; Nse O. Umoh; Francis O. Eko; Nalin Rastogi

SETTING Nigeria has the worlds fourth largest tuberculosis burden, and multidrug resistant tuberculosis (MDR-TB) represents a serious public health problem. OBJECTIVES To describe the demography of TB patients and determine the susceptibility of Mycobacterium tuberculosis isolates to the major TB drugs. METHODS One hundred and thirty-seven newly diagnosed TB patients (26 (19%) being HIV positive) from all age groups were recruited into the study. Each specimen was cultured using BACTEC MGIT960, followed by inoculation and growth on Lowenstein-Jensen (LJ) medium. Primary identification was carried out using an immunochromatographic technique (Capilia TB-Neo), and further confirmed by genotyping. Drug susceptibility testing (DST) was carried out by the agar proportion method. RESULTS Of the 97 pure mycobacterial cultures on LJ medium, 81 (83.5%) isolates were identified as M. tuberculosis complex, while 16 (16.5%) were Capilia negative. DST was carried out on 58 isolates. The drug susceptibility pattern showed that resistance occurred in 16 (27.6%) for streptomycin, 11 (19%) for isoniazid, 9 (16%) for rifampicin, and 10 (17.2%) for ethambutol. Rifampicin monoresistance occurred in 2 (3.4%) cases. MDR (combined resistance to isoniazid and rifampicin), also involving resistance to streptomycin and ethambutol, occurred in 6/58 (10.3%) isolates; although laboratory cross-contamination could not be excluded in 4/6 MDR strains with identical MIRU patterns characterized by consecutive strain numbers. Considering that first out of these 4 isolates was not due to laboratory carryover, the results of this study still report a minimal MDR-TB rate of 3/58 (5.2%) among newly diagnosed TB patients in Cross River State, Nigeria. CONCLUSIONS An increase in drug resistance was observed in this study as compared with previous studies in the country. Hence, introduction of culture in routine diagnostic mycobacteriology laboratories will prevent the emergence and dissemination of MDR-TB, while improved quality control strategies would in parallel prevent laboratory cross-contamination, thereby reducing mislabeling, unnecessary treatment, and drug toxicity for patients.


The International Journal of Mycobacteriology | 2013

Utility and diagnostic performance of Mycobacterium tuberculosis complex by two immunochromatographic assays as compared with the molecular Genotype assay in Nigeria

Benjamin Thumamo Pokam; Anne E. Asuquo; Khye Seng Goh; Lydia N. Abia-Bassey; Nalin Rastogi

Among the disadvantages of smear microscopy for detection of tuberculosis cases is its inability to differentiate between Mycobacterium tuberculosis (MTB) and non-tuberculous mycobacteria (NTM). This study evaluated two, new immunochromatographic assays - Capilia TB-Neo and SD Bioline - on unheated and heated cultures at 80°C for 30min respectively for their ability to discriminate between MTB complex and NTM as compared with the molecular Genotype assay. Mycobacteria used in the study were obtained from smear-positive specimens collected from patients at four major hospitals in Cross River State, Nigeria. Capilia TB-Neo and SD Bioline showed sensitivities of 98.8% and 93.8% respectively and 100% specificity for both assays. Heating the isolates did not significantly impact the test performance. Both tests are recommended for use in rapid differentiation of strains isolated in Nigeria.


Studies on Ethno-Medicine | 2009

Tuberculin Skin Test (TST) Indurations in Smear Positive TB Patients and Healthy Individuals in Calabar, Nigeria

Anne E. Asuquo; Adebayo N. Dairo; Lydia N. Abia-Bassey; Martin M. Meremiku; Benjamin P. Thumamo

Abstract The tuberculin skin test (TST) is used to detect latent Mycobacterium tuberculosis infection. WHO guidelines recommend 10mm as tuberculosis (TB) infection threshold for high risk individuals and 15mm for persons with no risk factors for TB. Nigeria is one of the 22 high burden countries for TB and her population is at risk of exposure to the TB germ. In an attempt to investigate indigenous TST indurations in a local population, 200 apparently healthy new students of tertiary institutions and other residents of Calabar - Nigeria metropolis that required the test for routine medical examination were examined. Each subject was injected intradermally with 0.1mL of 5TU of purified protein derivative (PPD) into the dorsal surface of the forearm. Indurated areas were measured after 48-72 hours of administration and results expressed in millimeters. 200 smear positive TB patients receiving treatment at the treatment center of the National TB Control program were used as the reference group. Results were statistically analysed using Chi square and T-test. Data obtained from the apparently healthy group show that approximately 31% of the individuals had indurations measuring 5-7mm while 29% were non-reactive. At least 25% of these individuals had indurations of >10mm compared to 95% of TB patients. Nine (4.5%) of TB patients have indurations <10mm. The results of this study imply that at least 25% of the healthy subjects are at risk of progressing to active disease when exposed to conditions that lower the individual’s immune status.


Journal of Medical Laboratory Science | 2007

Studies on Urine Specimens Adjudged Non-Significant for Bacteriuria

Anne E. Asuquo; B N Ekwere; S. J. Utsalo

The concept of significant bacteriuria continues to form the basis for the diagnosis of urinary tract infections in many laboratories, despite evidence that lower counts of bacteria may be associated with urinary tract infection (UTI). This study investigated the presence of pyuria, leucocyte esterase, nitrite and protein in urine specimens adjudged non-significant for bacteriuria. Methods used included urine centrifugation and microscopy for pyuria, and dip-stick reagent strip testing for leucocyte esterase, nitrites and protein. The presence of significant bacteriuria in the urine specimens was determined on Cysteine lactose electrolyte deficiency (CLED) agar and blood agar using the filter paper method. The cytological and chemical parameters diagnostic of UTI were found in the non-significant bacteriuric specimens as well as in the significant specimens; howbeit at a lower percentage. Similar bacteria were also isolated from both specimen types. Consequently, laboratory diagnosis of UTI on the basis of quantitative criteria should be reviewed since evidence abounds that lower bacterial counts may be observed in symptomatic women. Keywords : Urinary tract infection, non-significant bacteriuria, pyuria Journal of Medical Laboratory Sciences Vol. 14 (1) 2005: pp. 48-56


BMC Infectious Diseases | 2005

Bacterial isolates from blood cultures of children with suspected septicaemia in Calabar, Nigeria

Martin Meremikwu; Chukwuemeka E Nwachukwu; Anne E. Asuquo; Joseph Okebe; S. J. Utsalo


The International Journal of Mycobacteriology | 2015

A public-private partnership to reduce tuberculosis burden in Akwa Ibom State, Nigeria.

Anne E. Asuquo; Benjamin Thumamo Pokam; A. Adindu; E.O. Ibeneme; E. Ekpereonne; V. Obot

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K. L. Morgan

University of Liverpool

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Robert Kelly

University of Edinburgh

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Ian Handel

University of Edinburgh

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