PrimeStore MTM and OMNIgene Sputum for the Preservation of Sputum for Xpert MTB/RIF Testing in Nigeria

Background: Xpert MTB/RIF (GX) for tuberculosis (TB) diagnosis is often located in reference laboratories, and sputum needs to be transported using a cold chain. Transport media to preserve sputum are available, but performance data under programmatic conditions are limited. Methods: Sputum samples were collected from patients with presumptive TB in Nigeria. One sputum was transported in a cold chain, tested immediately with GX and cultured. One sputum was swabbed and stored in PrimeStore-Molecular-Transport-Medium (Primestore), and the remainder was stored in OMNIGene-sputum (Omnigene), kept for seven days and tested with GX. Results: Of 248 patients, 63 were fresh-sputum culture-positive and 56 GX-positive (sensitivity 88.9%, 95% CI: 78.4–95.4%). Four of 185 culture-negative patients were GX-positive (specificity 97.8%, 94.6–99.4%). Omnigene GX and Primestore GX were positive in 56/62 (90.3%, 80.1–96.4%) and 49/62 (79.0%, 66.8–88.3%) culture-positive, respectively, and 1/185 (99.5%, 97.0–100.0%) and 3/185 (98.4%, 95.3–99.7%) were culture-negative patients. 14 Human Immunodeficiency Virus (HIV)-infected and 44 HIV-uninfected patients were culture-positive. Omnigene and Primestore detected 12/14 (85.7%, 57.2–98.2%) and 5/14 (35.7%, 12.8–64.9%) HIV-infected and 41/44 (93.2%, 81.3–98.6%) HIV-uninfected culture-positive patients. Interpretation: Omnigene stored and fresh sputum samples had similar GX results. The GX results of Primestore-stored samples were similar to those found in the fresh sputum of non-HIV infected patients, but GX-positivity was lower in HIV-infected patients. This was likely due to the lower amount of bacilli collected by the swab and transferred to PrimeStore.

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PCR Method for Detection of Adenovirus in Urine of Healthy and Human Immunodeficiency Virus-Infected Individuals

Journal of Clinical Microbiology ◽

10.1128/jcm.36.11.3323-3326.1998 ◽

1998 ◽

Vol 36 (11) ◽

pp. 3323-3326 ◽

Cited By ~ 95

Author(s):

Marcela Echavarria ◽

Michael Forman ◽

John Ticehurst ◽

J. Stephen Dumler ◽

Patricia Charache

Keyword(s):

Human Immunodeficiency Virus ◽

Urine Samples ◽

Culture Positivity ◽

Immunodeficiency Virus ◽

History Of ◽

Pcr Method ◽

Urine Processing ◽

Culture Positive ◽

Culture Negative

Adenoviruses (AdV) cause diseases that range from localized, self-limited illnesses to fatal infections in immunocompromised patients. Culture is assumed to be sensitive but requires viable virus and up to 3 weeks for detection, and it can be inhibited by bacterial contamination. A new PCR method amplifying a region of the hexon gene was developed in order to detect AdV in urine more rapidly and with greater sensitivity than obtainable by culture technology. All 18 serotypes tested were detected. Quantitatively, with optimized urine processing, AdV PCR detected 0.2 PFU/ml (serotype 11) and 10 DNA copies/ml (serotype 2). Serially collected urine samples from human immunodeficiency virus (HIV)-infected patients with concurrent cytomegalovirus retinitis were divided into three groups: AdV culture-positive samples, AdV culture-negative or bacterially contaminated samples from patients with a history of AdV culture-positive urines, and AdV culture-negative samples from patients without a history of AdV culture positivity. Urine samples from healthy adults were also tested by culture and PCR to screen for asymptomatic shedding. Amplification was assessed with and without prior DNA purification. AdV was detected by PCR in 90% of culture-positive urines (100% of unclotted samples, e.g., those culture positive after storage for PCR testing), 71% of culture-negative or bacterially contaminated urines from AdV-infected patients, and 28% from AdV culture-negative patients. Healthy volunteers were culture negative for AdV, and 96% were PCR negative. The new AdV PCR method is rapid and sensitive and can detect viral DNA in samples for which culturing is problematic. The role of AdV replication during HIV infection merits further investigation with sensitive tools such as PCR.

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Culture-positive tuberculosis in human immunodeficiency virus type 1-infected children

The Pediatric Infectious Disease Journal ◽

10.1097/00006454-199807000-00005 ◽

1998 ◽

Vol 17 (7) ◽

pp. 599-604 ◽

Cited By ~ 33

Author(s):

H. SIMON SCHAAF ◽

AGNETA GELDENDUYS ◽

ROBERT P. GIE ◽

MARK F. COTTON

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Immunodeficiency Virus ◽

Culture Positive

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Fungal dysbiosis in cirrhosis

Gut ◽

10.1136/gutjnl-2016-313170 ◽

2017 ◽

Vol 67 (6) ◽

pp. 1146-1154 ◽

Cited By ~ 40

Author(s):

Jasmohan S Bajaj ◽

Eric J Liu ◽

Raffi Kheradman ◽

Andrew Fagan ◽

Douglas M Heuman ◽

...

Keyword(s):

Standard Of Care ◽

High Rate ◽

Control Groups ◽

Cross Sectional ◽

And Control ◽

Uninfected Culture ◽

Culture Positive ◽

Culture Negative

ObjectiveCirrhotics have a high rate of infections, which are increasingly fungal or culture-negative in nature. While infected cirrhotics have bacterial dysbiosis, the role of fungi is unclear. We aimed to evaluate gut bacterial and fungal dysbiosis in cross-sectional and longitudinal analyses of outpatient and inpatient cirrhotics and prediction of hospitalisations.MethodsCross-sectional: Age-matched controls, outpatients (with/without antibiotics) and hospitalised uninfected, culture-negative and culture-positive cirrhotics were included and followed for 90 days. Longitudinal: Three studies were conducted: (1) cirrhotics followed over 6 months, (2) outpatient cirrhotics administered antibiotics per standard of care for 5 days and (3) cirrhotics and controls administered omeprazole over 14 days. In all studies, stool bacterial/fungal profiles were analysed.ResultsCross-sectional: In 143 cirrhotics and 26 controls, bacterial and fungal diversities were significantly linked. Outpatients on antibiotics and patients with culture-positive infections had the lowest diversities. Bacterial and fungal correlations were complex in uninfected, outpatient and control groups but were markedly skewed in infected patients. 21% were admitted on 90-day follow-up. A lower Bacteroidetes/Ascomycota ratio was associated with lower hospitalisations. Longitudinal: Fungal and bacterial profiles were stable on follow-up (5 days and 6 months). After antibiotics, a significantly reduced bacterial and fungal diversity, higher Candida and lower autochthonous bacterial relative abundance were seen. After omeprazole, changes in bacterial diversity and composition were seen but fungal metrics remained stable.ConclusionThere is a significant fungal dysbiosis in cirrhosis, which changes differentially with antibiotics and proton pump inhibitor use, but is otherwise stable over time. A combined bacterial–fungal dysbiosis metric, Bacteroidetes/Ascomycota ratio, can independently predict 90-day hospitalisations in patients with cirrhosis.Clinical trial numberNCT01458990.

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A clinical specimen collection and transport medium for molecular diagnostic and genomic applications

Epidemiology and Infection ◽

10.1017/s0950268810002384 ◽

2010 ◽

Vol 139 (11) ◽

pp. 1764-1773 ◽

Cited By ~ 15

Author(s):

L. T. DAUM ◽

S. A. WORTHY ◽

K. C. YIM ◽

M. NOGUERAS ◽

R. F. SCHUMAN ◽

...

Keyword(s):

Nucleic Acid ◽

Real Time ◽

Genetic Characterization ◽

Clinical Specimen ◽

Molecular Transport ◽

Cold Chain ◽

Molecular Diagnostic ◽

Rt Pcr ◽

Transport Medium ◽

Specimen Collection

SUMMARYPathogen detection and genetic characterization has dramatically changed in recent years. Clinical laboratories are transitioning from traditional culture and primer-specific sequencing to more robust and rapid nucleic acid testing such as real-time PCR and meta-genomic characterization, respectively. Specimen collection is the first step in any downstream molecular diagnostic procedure. PrimeStore Molecular Transport Medium (MTM) is an optimized blend of nucleic acid stabilizing reagents that includes a non-specific internal positive control that can be amplified using real-time RT–PCR for tracking the integrity of a specimen from the point of collection to detection. PrimeStore MTM is shown here to effectively kill pathogens, including highly pathogenic H5 influenza virus, inactivate nucleases and to protect and preserve released RNA at ambient temperature for up to 30 days for downstream real-time and traditional RT–PCR detection and genetic characterization. PrimeStore MTM is also compatible with a variety of commercial extraction kits. PrimeStore is suited for routine clinical specimens and has added utility for field collection in remote areas, triage centres, border crossings and during pandemics where cold-chain, transport, and dissemination of potentially infectious pathogens are a concern.

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Dendritic Cells Transmit Human Immunodeficiency Virus Type 1 to Monocytes and Monocyte-Derived Macrophages

Journal of Virology ◽

10.1128/jvi.72.8.6671-6677.1998 ◽

1998 ◽

Vol 72 (8) ◽

pp. 6671-6677 ◽

Cited By ~ 33

Author(s):

Laco Kacani ◽

Ines Frank ◽

Martin Spruth ◽

Michael G. Schwendinger ◽

Brigitte Müllauer ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

T Cells ◽

Dendritic Cells ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Productive Infection ◽

Lower Amount ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT Previous studies have shown that human immunodeficiency virus type 1 (HIV-1) exploits dendritic cells (DC) to replicate and spread among CD4+ T cells. To explain the predominance of non-syncytium-inducing (NSI) over syncytium-inducing (SI) strains during the initial viremia of HIV, we investigated the ability of blood monocyte (Mo)-derived DC to transmit HIV-1 to CD4+ cells of the monocytoid lineage. First, we demonstrate that in our system, DC are able to transmit NSI strains, but not SI strains, of HIV-1 to fresh blood Mo and to Mo-derived macrophages (MDM). To establish a productive infection, a 10-fold-lower amount of virus was necessary for DC-mediated transmission of HIV-1 to Mo than in case of cell-free infection. Second, immature CD83− DC (imDC) transmit virus to Mo and MDM with higher efficacy compared to mature CD83+DC (maDC); this finding is in contrast to data previously obtained with CD4+ T cells. Third, maturation from imDC to maDC efficiently silenced expression of β2-integrins CD11b, CD11c, and CD18 by maDC. Moreover, monoclonal antibody against CD18 inhibited transmission of HIV-1 from imDC to Mo. We propose that the adhesion molecules of the CD11/CD18 family, involved in cell-cell interactions of DC with the microenvironment, may play a major role in imDC-mediated HIV-1 infection of Mo and MDM.

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The Value of Tests of Cure following Cervical Chlamydial Infection

International Journal of STD & AIDS ◽

10.1177/095646249300400102 ◽

1993 ◽

Vol 4 (1) ◽

pp. 5-7 ◽

Cited By ~ 4

Author(s):

DJ White ◽

CH Mann ◽

RS Matthews ◽

J G Leeming ◽

J C Clay

Keyword(s):

Chlamydial Infection ◽

Positive Culture ◽

Detection Methods ◽

Contact Tracing ◽

Chlamydia Infection ◽

Transport Medium ◽

Significant Difference ◽

Culture Positive ◽

Positive Results ◽

Culture Negative

Test of cure (TOC) was performed 2, 4 and 6 weeks after treatment for cervical chlamydia infection with 10–14 days of Deteclo one tablet twice daily, erythromycin 500 mg twice daily or doxycycline 100 mg twice daily. Testing was by chlamydia culture and IDEIA™ (DAKO diagnostics Ltd). Discrepant results were subsequently checked by immunofluorescence (Syva MicroTrak™) of both sets of left over transport media. Two hundred and three patients attended on at least one occasion; 189, 146 and 107 at 2, 4 and 6 weeks respectively. Of these 127, 70 and 34, respectively, denied sexual intercourse or had consistently used condoms. Fourteen were positive over the study period by either or both methods of detection. Of 8 culture positive results 3 were negative by IDEIA. Two of these had elementary bodies (EBs) on immunofluorescence of both sets of saved transport media. One had EBs on immunofluorescence of the saved culture transport medium only. None of the 6 IDEIA positive, culture negative patients had immunofluorescent EBs in the IDEIA transport media although one had EBs in the saved culture transport medium. One IDEIA suspicious, culture negative patient had EBs in both sets of saved transport media. There was no significant difference in the rate of chlamydia detection from patients admitting to or denying unprotected intercourse. TOC has a low yield in cases of cervical chlamydial infection when there has been careful contact tracing and treatment has been completed. If TOC is performed culture should be used if available and where antigen detection methods are used confirmation should be sought for any positive results.

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Treatment of chancroid with azithromycin

International Journal of STD & AIDS ◽

10.1258/0956462961917230 ◽

1996 ◽

Vol 7 (1_suppl) ◽

pp. 9-12 ◽

Cited By ~ 17

Author(s):

R C Ballard ◽

Htun Ye ◽

A Matta ◽

Y Dangor ◽

F Radebe

Keyword(s):

Single Dose ◽

South African ◽

Comparative Evaluation ◽

Benzathine Penicillin ◽

Ulcer Disease ◽

Genital Ulcer Disease ◽

Immunodeficiency Virus ◽

Cure Rates ◽

Culture Positive ◽

Culture Negative

A randomized, comparative study undertaken in Nairobi, Kenya and a non-comparative evaluation undertaken in Carletonville, South Africa have both shown that a single oral dose of azithromycin 1 g is effective in the treatment of the genital ulcer disease (GUD), chancroid, with cure rates of 89% and 92% recorded respectively. While treatment failure was associated with human immunodeficiency virus seropositivity and lack of circumcision in Kenya, no such association could be found in the South African study. In both series, azithromycin treatment resulted in cure of both Haemophilus ducreyi culture-positive and culture-negative cases of GUD, including two cases subsequently diagnosed as lymphogranuloma venereum. A combination of single-dose azithromycin with single-dose benzathine penicillin may provide effective ‘single-visit’ syndromic treatment for GUD in many developing countries.

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