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Survey and Visual Detection of Zaire ebolavirus in Clinical Samples Targeting the Nucleoprotein Gene in Sierra Leone.

Li H, Wang X, Liu W, Wei X, Lin W, Li E, Li P, Dong D, Cui L, Hu X, Li B, Ma Y, Zhao X, Liu C, Yuan J - Front Microbiol (2015)

Bottom Line: Ebola virus (EBOV) can lead to severe hemorrhagic fever with a high risk of death in humans and other primates.The RT-LAMP assay detected the NP target sequence with a limit of 4.56 copies/μL within 45 min under 61°C, a similar even or increase in sensitivity than that of real-time reverse transcription-polymerase chain reaction (RT-PCR).Additionally, all pseudoviral particles or non- Zaire EBOV genomes were negative for LAMP detection, indicating that the assay was highly specific for EBOV.

View Article: PubMed Central - PubMed

Affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.

ABSTRACT
Ebola virus (EBOV) can lead to severe hemorrhagic fever with a high risk of death in humans and other primates. To guide treatment and prevent spread of the viral infection, a rapid and sensitive detection method is required for clinical samples. Here, we described and evaluated a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method to detect Zaire ebolavirus using the nucleoprotein gene (NP) as a target sequence. Two different techniques were used, a calcein/Mn(2+) complex chromogenic method and real-time turbidity monitoring. The RT-LAMP assay detected the NP target sequence with a limit of 4.56 copies/μL within 45 min under 61°C, a similar even or increase in sensitivity than that of real-time reverse transcription-polymerase chain reaction (RT-PCR). Additionally, all pseudoviral particles or non- Zaire EBOV genomes were negative for LAMP detection, indicating that the assay was highly specific for EBOV. To appraise the availability of the RT-LAMP method for use in clinical diagnosis of EBOV, of 417 blood or swab samples collected from patients with clinically suspected infections in Sierra Leone, 307 were identified for RT-LAMP-based surveillance of EBOV. Therefore, the highly specific and sensitive RT-LAMP method allows the rapid detection of EBOV, and is a suitable tool for clinical screening, diagnosis, and primary quarantine purposes.

No MeSH data available.


Related in: MedlinePlus

Visual detection of RT-LAMP findings of 413 patients with clinically suspected infections in Sierra Leone. Sample information is listed in Supplemental Tables S1 and S2. A total of 307 positive (A) and 106 negative (B) samples were tested for EBOV infection by real-time RT-PCR and RT-LAMP. NC, negative control (distilled water); PC, positive control (artificial EBOV RNA).
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Figure 4: Visual detection of RT-LAMP findings of 413 patients with clinically suspected infections in Sierra Leone. Sample information is listed in Supplemental Tables S1 and S2. A total of 307 positive (A) and 106 negative (B) samples were tested for EBOV infection by real-time RT-PCR and RT-LAMP. NC, negative control (distilled water); PC, positive control (artificial EBOV RNA).

Mentions: The 417 clinical blood or swab samples were simultaneously analyzed by RT-LAMP and real-time RT-PCR. Of these, 307 patients were confirmed to be infected with EBOV, while 106 tested negative (Figure 4 and Table 2). A higher Ct value (Ct > 36) was recorded for the remaining four samples by RT-PCR, and green fluorescence was observed after ∼45 min by RT-LAMP, indicating a low level of EOBV RNA. We suggested that these patients should be monitored for 2 weeks in hospital.


Survey and Visual Detection of Zaire ebolavirus in Clinical Samples Targeting the Nucleoprotein Gene in Sierra Leone.

Li H, Wang X, Liu W, Wei X, Lin W, Li E, Li P, Dong D, Cui L, Hu X, Li B, Ma Y, Zhao X, Liu C, Yuan J - Front Microbiol (2015)

Visual detection of RT-LAMP findings of 413 patients with clinically suspected infections in Sierra Leone. Sample information is listed in Supplemental Tables S1 and S2. A total of 307 positive (A) and 106 negative (B) samples were tested for EBOV infection by real-time RT-PCR and RT-LAMP. NC, negative control (distilled water); PC, positive control (artificial EBOV RNA).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664619&req=5

Figure 4: Visual detection of RT-LAMP findings of 413 patients with clinically suspected infections in Sierra Leone. Sample information is listed in Supplemental Tables S1 and S2. A total of 307 positive (A) and 106 negative (B) samples were tested for EBOV infection by real-time RT-PCR and RT-LAMP. NC, negative control (distilled water); PC, positive control (artificial EBOV RNA).
Mentions: The 417 clinical blood or swab samples were simultaneously analyzed by RT-LAMP and real-time RT-PCR. Of these, 307 patients were confirmed to be infected with EBOV, while 106 tested negative (Figure 4 and Table 2). A higher Ct value (Ct > 36) was recorded for the remaining four samples by RT-PCR, and green fluorescence was observed after ∼45 min by RT-LAMP, indicating a low level of EOBV RNA. We suggested that these patients should be monitored for 2 weeks in hospital.

Bottom Line: Ebola virus (EBOV) can lead to severe hemorrhagic fever with a high risk of death in humans and other primates.The RT-LAMP assay detected the NP target sequence with a limit of 4.56 copies/μL within 45 min under 61°C, a similar even or increase in sensitivity than that of real-time reverse transcription-polymerase chain reaction (RT-PCR).Additionally, all pseudoviral particles or non- Zaire EBOV genomes were negative for LAMP detection, indicating that the assay was highly specific for EBOV.

View Article: PubMed Central - PubMed

Affiliation: Institute of Disease Control and Prevention, Academy of Military Medical Sciences Beijing, China.

ABSTRACT
Ebola virus (EBOV) can lead to severe hemorrhagic fever with a high risk of death in humans and other primates. To guide treatment and prevent spread of the viral infection, a rapid and sensitive detection method is required for clinical samples. Here, we described and evaluated a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method to detect Zaire ebolavirus using the nucleoprotein gene (NP) as a target sequence. Two different techniques were used, a calcein/Mn(2+) complex chromogenic method and real-time turbidity monitoring. The RT-LAMP assay detected the NP target sequence with a limit of 4.56 copies/μL within 45 min under 61°C, a similar even or increase in sensitivity than that of real-time reverse transcription-polymerase chain reaction (RT-PCR). Additionally, all pseudoviral particles or non- Zaire EBOV genomes were negative for LAMP detection, indicating that the assay was highly specific for EBOV. To appraise the availability of the RT-LAMP method for use in clinical diagnosis of EBOV, of 417 blood or swab samples collected from patients with clinically suspected infections in Sierra Leone, 307 were identified for RT-LAMP-based surveillance of EBOV. Therefore, the highly specific and sensitive RT-LAMP method allows the rapid detection of EBOV, and is a suitable tool for clinical screening, diagnosis, and primary quarantine purposes.

No MeSH data available.


Related in: MedlinePlus