Molecular Detection of Plasmodium knowlesi in a Dutch Traveler by Real-Time PCR

The new BD Max GC real-time PCR assay showed high clinical and analytical sensitivity and specificity. It can be an effective and accurate supplementary test for the BD ProbeTec GC Qx amplified DNA assay, which had suboptimal specificity, and might also be used for initial detection ofNeisseria gonorrhoeae.

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Molecular detection of Pneumocystis jirovecii by real-time PCR: Sensitivity depends on the target selection

International Journal of Infectious Diseases ◽

10.1016/j.ijid.2014.03.1188 ◽

2014 ◽

Vol 21 ◽

pp. 372 ◽

Cited By ~ 1

Author(s):

P. Jayaratne ◽

L. Dalle Vedove ◽

D. Yamamura

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Molecular Detection ◽

Target Selection ◽

Pneumocystis Jirovecii

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Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia

10.21203/rs.3.rs-30644/v3 ◽

2020 ◽

Author(s):

Nor Afizah Nuin ◽

Angelica Fiona Tan ◽

Yao Long Lew ◽

Kim A Piera ◽

Timothy William ◽

...

Keyword(s):

Diagnostic Accuracy ◽

Real Time ◽

Real Time Pcr ◽

Limit Of Detection ◽

Plasmodium Knowlesi ◽

Plasmodium Species ◽

Detection Methods ◽

Clinical Samples ◽

Public Health Issue ◽

Gene Target

Abstract Background : The monkey parasite Plasmodium knowlesi is an emerging public health issue in Southeast Asia. In Sabah, Malaysia, P. knowlesi is now the dominant cause of human malaria. Molecular detection methods for P. knowlesi are essential for accurate diagnosis and in monitoring progress towards malaria elimination of other Plasmodium species. However, recent commercially available PCR malaria kits have unpublished P. knowlesi gene targets or have not been evaluated against clinical samples. Methods : Two real-time PCR methods currently used in Sabah for confirmatory malaria diagnosis and surveillance reporting were evaluated: the QuantiFast™ Multiplex PCR kit (Qiagen, Germany) targeting the P. knowlesi 18S SSU rRNA; and the abTES™ Malaria 5 qPCR II kit (AITbiotech, Singapore), with an undisclosed P. knowlesi gene target. Diagnostic accuracy was evaluated using 52 P. knowlesi, 25 P. vivax , 21 P. falciparum , and 10 P. malariae clinical isolates, and 26 malaria negative controls, and compared against a validated reference nested PCR assay. The limit of detection (LOD) for each PCR method and Plasmodium species was also evaluated. Results : The sensitivity of the QuantiFast™ and abTES™ assays for detecting P. knowlesi was comparable at 98.1% (95%CI 89.7-100) and 100% (95%CI 93.2-100) respectively. Specificity of the QuantiFast™ and abTES™ for P. knowlesi was high at 98.8% (95%CI 93.4-100) for both assays. The QuantiFast™ assay demonstrated falsely-positive mixed Plasmodium species at low parasitaemias in both the primary and LOD analysis. Diagnostic accuracy of both PCR kits for detecting P. vivax , P. falciparum , and P. malariae was comparable to P. knowlesi . The abTES™ assay demonstrated a lower LOD for P. knowlesi of ≤0.125 parasites/µL compared to QuantiFast™ with a LOD of 20 parasites/µL. Hospital microscopy demonstrated a sensitivity of 78.8% (95%CI 65.3-88.9) and specificity of 80.4% (95%CI 67.6-89.8) compared to reference PCR for detecting P. knowlesi . Conclusion : The QuantiFast™ and abTES™ commercial PCR kits performed well for the accurate detection of P. knowlesi infections. Although the QuantiFast™ kit is cheaper, the abTES™ kit demonstrated a lower LOD, supporting its use as a second-line referral-laboratory diagnostic tool in Sabah, Malaysia.

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Serologic and Molecular Detection of Mycoplasma gallisepticum in Layer Flocks in South Marmara Region of Turkey

Journal of Advances in Microbiology ◽

10.9734/jamb/2021/v21i1030390 ◽

2021 ◽

pp. 29-37

Author(s):

Elçin Günaydın ◽

Özlem Kardoğan ◽

Gülşen Goncagül ◽

Yavuz Çokal

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Molecular Detection ◽

Preventive Measures ◽

Mycoplasma Gallisepticum ◽

Marmara Region ◽

Time Loss ◽

Serological Tests ◽

Primary Screening ◽

Pooled Samples

Background: Due to the economic impacts of Mycoplasma gallisepticum (MG) infection in poultry, it is essential to have a fast, reliable and accurate diagnostic test to diagnose the infection. Aims: It was aimed to examine the presence of MG in the South Marmara Region of Turkey where extensive commercial layer flocks exist by RPA, ELISA and real-time PCR. Materials and Methods: In the study, 981 sera and 160 tracheal swab samples (20 swabs per each flock) obtained from eight layer flocks were examined for the presence of MG-antibody by RPA, ELISA, and the presence of MG by real-time PCR, respectively. Results: MG-seropositive flock rate was determined to be 100% by RPA. Twenty-three of the RPA positive sera in each flock LA, LB, LC, LD, LF, LG, and 17 RPA positive sera in flock LE (due to 17 positive RPA sera obtained) were examined for the presence of MG antibody by ELISA, and MG-seropositive flock rate was determined to be 87.5%. As a result of the examination of a total of 32 tracheal swab samples (20 swabs perflock/5 swabs=4 pooled samples, 8 flocksX4 pooled samples= 32 samples) for the presence of MG, real-time PCR positive flock rate was found to be 75%. Conclusion: To decide the flock whether it is infected or not and the initiate effective preventive measures against MG infection as soon as possible; serology should be applied simultaneously with bacteriology and/or PCR to prevent time loss due to shortcomings of serological tests used as primary screening test such as cross reactions, sensitivity and specificity problems.

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