A multiplex polymerase chain reaction assay for genus-, group- and species-specific detection of mycobacteria

Kurabachew, Mekonnen; Enger, Øivind; Sandaa, Ruth‐Anne; Skuce, Robin; Bjorvatn, Bjarne

doi:10.1016/j.diagmicrobio.2004.03.010

Cited by 19 publications

(14 citation statements)

References 22 publications

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“…The diagnosis of TBLN has been a true challenge solely by clinical evidence in the developing countries due to the limited diagnostic facility on hand [18]. If the diagnosis of TBLN can be established faster, and the diagnostic process becomes less cumbersome for the patient, PCR may reduce a delay both in the diagnosis and in the start of treatment [19].…”

Section: Discussionmentioning

confidence: 99%

Concentration of fine needle aspirates similar to molecular method improves sensitivity of the diagnosis of tuberculous lymphadenitis in Addis Ababa, Ethiopia

et al. 2017

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BackgroundTuberculous lymphadenitis (TBLN) diagnosis has been a true challenge solely by clinical evidence in developing countries, due to limited the diagnostic facility on hand. However, the availability and affordability of available diagnostic tools in resource-limited settings like Ethiopia necessitates the quest for other techniques with added value over direct Z-N microscopy. Therefore, we aimed at to assess whether the concentration of lymph node aspirate similarly improves the detection rate of tuberculous lymphadenitis or not.Materials and methodsA cross-sectional study design was conducted on 132 individual subjects presumptive for tuberculous lymphadenitis from February to October 2013 in Addis Ababa, Ethiopia. Fine needle aspirate (FNA) samples were collected from the cases and cultured on Löwenstein-Jensen (LJ) slants. Identification of species and strains of mycobacteria was made by region of difference (RD) based polymerase chain reaction (PCR). Data entry and statistical analyses were performed by SPSS version 20. The confidence level of 95% was used for statistical significance.ResultA total of 132 study subjects were included in our study. Of these 56.1% (74/132) were positive for M. tuberculosis on culture. The detection rate of direct smear microscopy and the concentration method were 29.5 and 65.2% respectively. The sensitivity of direct smear microscopy was 43.2%, for concentrated smear microscopy 94.5%, for PCR 93.2% and for cytomorphology 95.4%. The level of agreement of concentrated ZN smear microscopy was 0.62 which was very similar with kappa of 0.58 of molecular (PCR) technique. AFB positivity by the concentration method and molecular method was increased in caseous aspirates as compared to purulent and hemorrhagic aspirates though it was not statistically significant (p-value = 0.18) and (p = 0.62) respectively.ConclusionThe concentration of FNA (Fine Needle Aspirate) aspirates for acid-fast smear microscopy similarly improves the sensitivity of acid fast bacilli in diagnosing of TBLN.

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Section: Discussionmentioning

confidence: 99%

Concentration of fine needle aspirates similar to molecular method improves sensitivity of the diagnosis of tuberculous lymphadenitis in Addis Ababa, Ethiopia

et al. 2017

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“…26,31 Sensitivity of the card test was also lower than suggested by previous literature 26 when estimated relative to qPCR. PCR frequently are considered to have high diagnostic Sp, 7,13,22,24 suggesting that it is a relatively good gold standard positive assay but can have less than perfect Se. When a test with almost perfect Sp but less than perfect Se (e.g., qPCR) is used as the reference standard, there will be some truly infected animals included in the noninfected group, but all noninfected animals will be correctly classified.…”

Section: Discussionmentioning

confidence: 99%

Diagnostic Accuracy of Methods for Detecting Anaplasma Marginale Infection in Lactating Dairy Cattle of Puerto Rico

et al. 2010

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Abstract. Bovine anaplasmosis (BA) is a hemoparasitic disease of great importance in cattle within the tropical and subtropical regions of the world. Control programs for BA require accurate diagnostic assays but validation can be challenging because the true disease status of all animals is frequently not known with certainty. The objective of this study was to estimate the accuracy of assays for detection of Anaplasma marginale infection in lactating dairy cattle of Puerto Rico using Bayesian methods without a perfect reference test. There were 2,331 cattle with complete diagnostic results sampled from 79 herds, and the prevalence of BA was estimated as 22% (95% probability interval [PI]: 19-25%). The sensitivity (Se) and specificity (Sp) of a major surface protein 5 competitive enzyme-linked immunosorbent assay (MSP-5 cELISA) were estimated as 99% (95% PI: 96-100%) and 89% (95% PI: 87-92%), respectively. The Se and Sp of a quantitative polymerase chain reaction (qPCR) were 67% (95% PI: 60-74%) and 99% (95% PI: 99-100%). The Se and Sp of a card agglutination test were 34% (95% PI: 29-39%) and 99% (95% PI: 99-100%). Area under the receiveroperating characteristic curve for the MSP-5 cELISA was 0.748 (95% PI: 0.71-0.79). The MSP-5 cELISA appears to be the test of choice for screening cattle for subclinical BA based on the high estimated Se, rapidity of results, relative low cost, and ease of standardization.

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“…1998) have been exploited for the differentiation of the two species. Kurabachew et al . (2004) developed a multiplex PCR assay targeting 23 s rDNA and the oxyR gene to distinguish between M. tuberculosis and M. bovis , where M. tuberculosis strains produce both the 23 s rDNA and the oxyR amplicons, whereas only the 23 s rDNA amplicons are produced by M. bovis .…”

Section: Molecular Diagnosticsmentioning

confidence: 99%

Mycobacterium bovis: the importance of milk and dairy products as a cause of human tuberculosis in the UK. A review of taxonomy and culture methods, with particular reference to artisanal cheeses

Rowe

Donaghy

2008

Int J of Dairy Tech

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A multiplex polymerase chain reaction assay for genus-, group- and species-specific detection of mycobacteria

Cited by 19 publications

References 22 publications

Concentration of fine needle aspirates similar to molecular method improves sensitivity of the diagnosis of tuberculous lymphadenitis in Addis Ababa, Ethiopia

Concentration of fine needle aspirates similar to molecular method improves sensitivity of the diagnosis of tuberculous lymphadenitis in Addis Ababa, Ethiopia

Diagnostic Accuracy of Methods for Detecting Anaplasma Marginale Infection in Lactating Dairy Cattle of Puerto Rico

Mycobacterium bovis: the importance of milk and dairy products as a cause of human tuberculosis in the UK. A review of taxonomy and culture methods, with particular reference to artisanal cheeses

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