Clinical Diagnostic Microbiology |

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Clinical Diagnostic Microbiology

Research Article

Pages: 1 - 4

Diagnosis of Pulmonary Tuberculosis Using Genotype MTBDRplus Assay in Three Local Government Primary Health Centres of Osun State, Nigeria- a Pilot Study

Aderemi Oludiran Kehinde and Adetayo Emmanue Adetoye


DOI: 10.4172/2161-0703.S3-001

Study background: Tuberculosis including multi-drug resistant form is not adequately diagnosed in spite of the fact that it constitutes a major public health challenge. This study was carried out to obtain data on laboratory diagnosis of pulmonary tuberculosis at the three local government areas in Osun state, Nigeria.

Methods: Six month pilot study was carried out at Iwo, Irewole and Ede North LGAs, all located in Osun state, Nigeria. Socio-demographic and clinical information of the subjects were obtained using a pre-tested questionnaire. Sputum samples were collected from symptomatic pulmonary tuberculosis patients, stained with Zeihl-Neelsen (ZN) reagents and cultured on egg-based Lowenstein-Jensen medium. The medium was incubated at 37°C for eight weeks. Acid Fast Bacilli was confirmed by repeat Z-N staining. Drug resistance testing of the isolates was done using Genotype MTBDRplus. Serum was screened for HIV test using recombinant ELISA. Those that were screened positive were retested using Capillus HIV 1 and 2.   Results: Of the 75 subjects studied, seven (9.3%) were over 60 years while the majority, 45 (76.0%) were aged 21-60 years. Thirty-four (45.3%) had their sputum positive for acid fast bacilli (AFB) while 24 (32.0%) were culture positive. Among the 24 isolates, 6 were identified to be non tuberculos mycobacteria (NTM) by molecular methods. The HIV prevalence rate was observed to be 18.6% among all the study participants. Six out of 34 AFB positive persons (17.6%) were also positive for HIV specific antibody, while eight (19.5%) of the AFB negative persons were positive for HIV. There is no significant difference in the incidence of TB between the HIV infected and uninfected groups (p>0.05).Among the 18 mycobacterial isolates (excluding NTM), one strain (5.5%) was found to be resistant to rifampicin and isoniazid (Multi-Drug Resistant-MDR) while three were resistant to isoniazid alone..   Conclusion: This pilot study reveals the existence of MDR-TB and warrants well designed studies to ascertain the magnitude of the problem.

Research Article

Pages: 0 - 0

The Spreading of Flaviviruses over the Continental Divide: a Challenge for Serologic Diagnostics

Sissy Therese Sonnleitner, Josef Simeoni, Raphaela Baumgartner, Roland Zelger, Angelika Prader, Grazia Piccolin, Norbert Nowotny and Gernot Walder


DOI: 10.4172/2161-0703.S3-002

West Nile virus and Usutu virus are spreading over Europe. The last confirmed outbreaks of West Nile virus took place in Greece in 2011 and Northern Italy, 2012. A serious outbreak of Usutu virus in birds occurred in Germany in 2010 and 2011, indicating an increased risk of Usutu virus infections in humans. Sera of 1,607 healthy blood donors in the Tyrols (Austria and Italy) were investigated for the presence of IgG antibodies for tick-borne encephalitis virus and West Nile virus by ELISA as well as for Usutu virus by in-house immunofluorescence assay. Furthermore, 2,571 mosquitoes were trapped in the study area and screened for flaviviral and West Nile-specific RNA by RT-PCR. Initial results indicated seroprevalence rates of 46.2% for West Nile virus in North Tyrol and 0.5% in South Tyrol as well as 16.5% and 0.3% for Usutu virus, respectively, which turned out to be false positive cross-reactions with antibodies against tick-borne encephalitis virus or other vaccine-associated flaviviral antibodies by adjacent neutralization assays. Actually, neutralizing antibodies against West Nile virus were found in five sera (0.5% seroprevalence), against Usutu virus in one serum (0.1% seroprevalence). This study shows that the Tyrolean population does not come into immunoreactive contact with West Nile virus or Usutu virus and that no flavivirus could be detected in trapped mosquitoes. Furthermore, the study provides cross-reactivities of up to 58.8% (460 of 782) of flaviviral antibodies in ELISA-testing and up to 21.5% (168 of 782) in immunofluorescence assay and that cross-reactivity increases with increasing number of vaccinations against Flaviviruses. This study illuminates the degree of cross-reactivities of flaviviral antibodies in a healthy blood donor collective and shows the difficulties in the differentiation of flaviviral antibodies in serologic diagnostics.

Research Article

Pages: 0 - 0

Clinical Validation of the BD Onclarity├?┬ó├?Ô??├?┬ó HPV Assay Using a Non-Inferiority Test

Ditte Møller Ejegod, Jesper Bonde, Itziar Serrano, Kate S Cuschieri, William A Nussbaumer, Laurence M Vaughan, Amar S Ahmad and Jack Cuzick


DOI: 10.4172/2161-0703.S3-003

The clinical performance of The BD Onclarity™ HPV Assay, a novel type-specific real-time E6/E7-based PCR assay, was evaluated for clinical and analytical performance using the Meijer et al. international guidelines for validation of high-risk HPV tests. Assay performance was found to be similar to the reference method, Hybrid Capture 2 (HC2, QIAGEN) using PreservCyt® Specimens (Hologic®, Inc.). In addition, the fully automated assay was found to have excellent intra- and inter-laboratory reproducibility (98.6% (kappa=0.967) and 98.4% (kappa=0.962), respectively). These data show that the BD Onclarity™ HPV Assay fulfills the clinical validation requirements for a HPV based cervical cancer screening assay (Meijer et al. International journal of cancer 2009; 124: 516-520.).

Research Article

Pages: 0 - 0

Hospital-based Sentinel Surveillance of Haemophilus Influenzae Type B among Children in Burkina Faso, 2004-2012: Impact of Vaccine Introduction

Idrissa Sanou, Isidore Juste O Bonkoungou, Isabelle Bicaba, Ali Ouedraogo, Fabienne Soudre, Sylvain Zeba, Isaïe Medah, Ludovic Kam and Lassana Sangare


DOI: 10.4172/2161-0703.S3-004

Background: Haemophilus influenzae type b (Hib) is a leading cause of childhood bacterial meningitis in Africa. This study assessed the impact of Hib conjugate vaccine introduced in 2006 into the Expanded Program on Immunization (EPI) in Burkina Faso.

Methods: From 2004-2012, we conducted hospital-based surveillance for invasive Hib disease among children <5 years of age with suspected bacterial meningitis. All cerebrospinal fluid (CSF) samples were tested using culture methods and/or PCR. Incidences calculated using population denominators were compared between pre-vaccine (2004-2005) and post- vaccine (2007-2012) periods.

Results: 3928 cases of suspected meningitis were identified from the pediatric service, 231(5.9%) of whom had a bacterial pathogen confirmed. Hib was found in 80 (34.6%) of confirmed cases, followed by Streptococcus pneumoniae in 76 (32.90%) and Neisseria meningitidis in 54 (23.38%). The average annual incidence (per 1,00,000 children) of Hib meningitis was 4.11; the annual incidence of Hib meningitis declined by 94.13% from 16 per 1,00,000 in pre-vaccine (2004-2005) to 0.94 per 100 000 in post-vaccine (2007-2012) periods. None of the 80 Hib confirmed cases was immunized and no death was notified after the introduction of the vaccine.

Conclusion: Admissions for Hib meningitis in the department of pediatrics at University Hospital, CHU-Yalgado Ouédraogo have practically disappeared two years after the introduction of the Hib vaccine into Burkina Faso’s Expanded Program on Immunization.

Review Article

Pages: 0 - 0

Probiotic Properties of Lactic Acid Bacteria from Human Milk

Desalegn Amenu


DOI: 10.4172/2161-0703.S3-005

Probiotics are preparation of live microorganisms that have beneficial effects on the health of their host. Although probiotic strains can be isolated from many sources, for human applications, the main criteria are that it must be from human origin. Milk is an important nutrient source for neonates. Previous studies have shown that human breast milk has beneficial effects on the health of neonates. One reason for these beneficial effects might be explained by the micro flora of human breast milk including beneficial lactic acid-producing bacteria. The main aim of this paper is to review best practices in isolation and characterization of lactic acid bacteria from human milk by biochemical and molecular methods and also determine the probiotic properties of lactic acid-producing bacteria. Most lactic acidproducing bacteria have effective antimicrobial activity against food-borne pathogens like Salmonella, thyphimurium CCM 5445, Escherichia coli O157:H7 NCTC 129000 and Escherichia coli NRRL B-3008. Data show that these isolates possess probiotic properties, as demonstrated by using amplification and restriction fragment length polymorphism (RFLP) of 16S ribosomal DNA (rDNA) and 16S sequencing. Therefore, it is observed that, human milk is a source of potential probiotic strains.

Mini Review

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The Role of Novel Molecular Techniques for Tuberculosis Diagnostics in the WHO European Region

Soudeh Ehsani, Martin van den Boom, Christopher Gilpin and Masoud Dara


DOI: 10.4172/2161-0703.S3-006

Tuberculosis and particularly multidrug resistant TB remains a major public health concern in the WHO European Region. In order to reduce the burden of the disease, early, accurate and rapid diagnoses play a crucial role. Sputum smear microscopy that has long been used as the main TB diagnostic method detects TB only with low sensitivity and phenotypic culture-based methods require several weeks and high biosafety laboratory infrastructures. The use of the WHO endorsed rapid molecular method for simultaneous detection of TB and rifampicin resistance detection has promising advantages to overcome these challenges and lead to major advances for early and accurate diagnosis of TB. With better understanding appearing discordances between phenotypic and genotypic methods, as well as the development and implementation of regionally adapted diagnostic algorithms, the precision of result acquisition and interpretation can advance even further.

Case Report

Pages: 0 - 0

Acute Blindness in a Child after Quinine treatment for Severe Malaria

Verner N. Orish and Ilechie A Alex


DOI: 10.4172/2161-0703.S3-007

Although blurred vision from quinine toxicity is well documented in literature, an acute effect leading to blindness is infrequently reported. We report a case of sudden bilateral blindness in a child after quinine treatment for severe malaria. Ocular examination revealed no acute pathology except widely dilated non-reactive pupils. The site within the retina of the toxic effect of quinine remains controversial. Literature on proposed mechanisms is reviewed in this article. This case highlights the potential toxic effects of quinine in children and the lack of a characteristic fundi appearance.

Research Article

Pages: 0 - 0

Effect of Argan Oil-Hydrogen Peroxide Mixture on Mycobacterium tuberculosis- In Vitro

Anwar K. AL-Saffar and Hussein O. M. Al-Dahmoshi


DOI: 10.4172/2161-0703.S3-008

Objective: Tuberculosis (TB) is the threatful infectious disease that dwell about one third of worlds. Among Middle East, Iraq occupy the fifth order country endemic with TB.

Methodology: Six mixtures of Argan oil: 1.5% H2O2 were prepared at different ratio: 1=0.75:9.25, 2=1:9, 3=1.5:8.5, 4=2:8, 5=2.5:7.5, 6=3:7 and mixed with Lowenstein-Jensen (L-J) medium. Seventeen Mycobacterium tuberculosis isolates got from the consultants clinic of respiratory and thoracic diseases in Hilla city-Iraq. All isolates cultured on LJ medium containing the mixture of Argan oil: 1.5% H2O2 in different ratio and control tube (LJ without mixture) also used. The growth monitored per a day and recorded at different interval and the inhibition (no growth) were recorded.

Results: The effect of Argan oil: 1.5% H2O2 mixture on the Mycobacterium tuberculosis were investigated over three incubation intervals (14th -20th, 21st -28th and after 29th day). The results revealed the high Argan concentration mixture 2.5:7.5 and 3:7 give excellent inhibitory effect on Mycobacterium tuberculosis during full periods of incubation with inhibition percentage (64.71%-82.35%).

Conclusion: The current study aims to investigate the effects of argan oil on Mycobacterium tuberculosis and conclude the anti-mycobacterial activity of argan oil as a safe medication for prophylaxis and treatment of TB.

Research Article

Pages: 0 - 0

Economic Impact of Microarray-Based Rapid Detection of Gram-Positive Organisms in Blood Cultures

Paul Mariani, Jesse Breidenbach and Felix Roth


DOI: 10.4172/2161-0703.S3-009

We assessed the clinical and economic impact of using the microarray-based Verigene Gram positive blood culture (BC-GP) test to identify gram positive bacteria in blood cultures. The main advantage of employing this test in comparison to conventional culture techniques resided on identifying the causative organism an average of 42.8 hours prior than conventional culture techniques. The microarray-based test results allowed providers to tailor antibiotic therapy and decide whether to admit/discharge a patient sooner, which reflected over $21,000 in hospital savings for the 105 cases analyzed in this study. The implementation of the Verigene BC-GP in our Microbiology Laboratory presented an economic advantage when compared to conventional workup and was shown to improve ongoing antimicrobial stewardship and patient care.

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Citations: 1059

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