Detection of Circulating Aspergillus fumigatus DNA by Real-Time PCR Assay of Large Serum Volumes Improves Early Diagnosis of Invasive Aspergillosis in High-Risk Adult Patients under Hematologic Surveillance

ABSTRACT We evaluated extended-interval dosing of the investigational echinocandin rezafungin (1, 4, and 16 mg/kg on days 1, 4, and 7 postinoculation) for the treatment of disseminated invasive aspergillosis caused by azole-resistant Aspergillus fumigatus. Survival was significantly improved in mice treated with each dose of rezafungin and supratherapeutic posaconazole (20 mg/kg twice daily). Kidney fungal burden, as measured by quantitative real-time PCR, was also significantly reduced in mice treated with rezafungin although variability was observed.

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Development and evaluation of a rapid direct DNA extraction and real-time PCR assay for early detection of methicillin resistant Staphylococcus aureus (MRSA) in high-risk patients

Journal of Infection ◽

10.1016/j.jinf.2007.04.077 ◽

2007 ◽

Vol 55 (3) ◽

pp. e63

Author(s):

Lucy Elston ◽

TatShing Yam ◽

Peter Marsh

Keyword(s):

Staphylococcus Aureus ◽

High Risk ◽

Early Detection ◽

Real Time ◽

Dna Extraction ◽

Real Time Pcr ◽

Methicillin Resistant Staphylococcus Aureus ◽

Pcr Assay ◽

Risk Patients ◽

Direct Dna Extraction

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Roles of the multiplex real-time PCR assay and β-D-glucan in a high-risk population for intra-abdominal candidiasis (IAC)

Medical Mycology ◽

10.1093/mmy/myz123 ◽

2019 ◽

Vol 58 (6) ◽

pp. 789-796 ◽

Cited By ~ 1

Author(s):

J Fortún ◽

M J Buitrago ◽

F Gioia ◽

E Gómez-Gª de la Pedrosa ◽

M E Alvarez ◽

...

Keyword(s):

High Risk ◽

Real Time ◽

Real Time Pcr ◽

Volume Overload ◽

Diagnostic Methods ◽

Practice Volume ◽

High Risk Population ◽

Pcr Assay ◽

European Consensus ◽

Abdominal Candidiasis

Abstract Multiplex quantitative real-time PCR (MRT-PCR) using blood can improve the diagnosis of intra-abdominal candidiasis (IAC). We prospectively studied 39 patients with suspected IAC in the absence of previous antifungal therapy. Blood cultures, MRT-PCR, and β-D-glucan (BDG) in serum were performed in all patients. IAC was defined according to the 2013 European Consensus criteria. For MRT-PCR, the probes targeted the ITS1 or ITS2 regions of ribosomal DNA. Candidaemia was confirmed only in four patients (10%), and IAC criteria were present in 17 patients (43.6%). The sensitivity of MRT-PCR was 25% but increased to 63.6% (P = .06) in plasma obtained prior to volume overload and transfusion; specificity was above 85% in all cases. BDG performance was improved using a cutoff > 260 pg/ml, and improvement was not observed in samples obtained before transfusion. In this cohort of high risk of IAC and low rate of bloodstream infection, the performance of non-culture-based methods (MRT-PCR or BDG) was moderate but may be a complementary tool given the limitations of diagnostic methods available in clinical practice. Volume overload requirements, in combination with other factors, decrease the accuracy of MRT-PCR in patients with IAC.

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Clinical Validation of the HPV-Risk Assay, a Novel Real-Time PCR Assay for Detection of High-Risk Human Papillomavirus DNA by Targeting the E7 Region

Journal of Clinical Microbiology ◽

10.1128/jcm.03195-13 ◽

2014 ◽

Vol 52 (3) ◽

pp. 890-896 ◽

Cited By ~ 33

Author(s):

A. T. Hesselink ◽

J. Berkhof ◽

M. L. van der Salm ◽

A. P. van Splunter ◽

T. H. Geelen ◽

...

Keyword(s):

Human Papillomavirus ◽

High Risk ◽

Real Time ◽

Real Time Pcr ◽

Clinical Validation ◽

Pcr Assay ◽

Papillomavirus Dna

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Development and validation of a quantitative real-time PCR assay for the early diagnosis of coccidioidomycosis

Diagnostic Microbiology and Infectious Disease ◽

10.1016/j.diagmicrobio.2014.01.029 ◽

2014 ◽

Vol 79 (2) ◽

pp. 214-221 ◽

Cited By ~ 18

Author(s):

Sara Gago ◽

María José Buitrago ◽

Karl V. Clemons ◽

Manuel Cuenca-Estrella ◽

Laurence F. Mirels ◽

...

Keyword(s):

Early Diagnosis ◽

Real Time ◽

Real Time Pcr ◽

Quantitative Real Time Pcr ◽

Pcr Assay ◽

Development And Validation

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Use of Multiplex Real-Time PCR To Diagnose Scrub Typhus

Journal of Clinical Microbiology ◽

10.1128/jcm.02181-16 ◽

2017 ◽

Vol 55 (5) ◽

pp. 1377-1387 ◽

Cited By ~ 12

Author(s):

Wiwit Tantibhedhyangkul ◽

Ekkarat Wongsawat ◽

Saowaluk Silpasakorn ◽

Duangdao Waywa ◽

Nuttawut Saenyasiri ◽

...

Keyword(s):

Early Diagnosis ◽

Real Time ◽

Real Time Pcr ◽

Scrub Typhus ◽

Febrile Illness ◽

Asia Pacific ◽

Diagnostic Tools ◽

Pcr Assay ◽

Content Type ◽

Serologic Tests

ABSTRACTScrub typhus, caused byOrientia tsutsugamushi, is a common cause of acute undifferentiated febrile illness in the Asia-Pacific region. However, its nonspecific clinical manifestation often prevents early diagnosis. We propose the use of PCR and serologic tests as diagnostic tools. Here, we developed a multiplex real-time PCR assay using hydrolysis (TaqMan) probes targetingO. tsutsugamushi47-kDa,groEL, and human interferon beta (IFN-β gene) genes to improve early diagnosis of scrub typhus. The amplification efficiency was higher than 94%, and the lower detection limit was 10 copies per reaction. We used a human gene as an internal DNA quality and quantity control. To determine the sensitivity of this PCR assay, we selected patients with confirmed scrub typhus who exhibited a clear 4-fold increase in the level of IgG and/or IgM. The PCR assay result was positive in 45 of 52 patients, indicating a sensitivity of 86.5% (95% confidence interval [CI]: 74.2 to 94.4). The PCR assessment was negative for all 136 non-scrub typhus patients, indicating a specificity of 100% (95% CI: 97.3 to 100). In addition, this test helped diagnose patients with inconclusive immunofluorescence assay (IFA) results and using single blood samples. In conclusion, the real-time PCR assay proposed here is sensitive and specific in diagnosing scrub typhus. Combining PCR and serologic tests will improve the diagnosis of scrub typhus among patients presenting with acute febrile illness.

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