Development and evaluation of a reverse transcription loop-mediated isothermal amplification assay for rapid detection of a new SFTS bunyavirus

Yang, Guoliang; Li, Bing; Liu, Lanzheng; Wei-chu, Huang; Zhang, Wen; Liu, Yuandong

doi:10.1007/s00705-012-1348-1

Cited by 32 publications

(31 citation statements)

References 14 publications

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“…When combined with a fluorescent detection reagent, visible results could be determined by showing a color change within 30 min. 26 Another similar assay, which incorporates reverse transcription-cross-priming amplification with a vertical flow visualization strip based on the M segment of the SFTSV, could complete the entire procedure from specimen processing to result reporting within 2 h. 27 As for serodiagnosis, the in-house Mac-ELISA assay, indirect ELISA assay, and double antigen sandwich ELISA assay have been established and optimized to test virus specific IgM, IgG and total antibodies in serum samples of patients, respectively. 1,14 Indirect immunofluorescence assays were also established to test the virus specific antibodies using previously fixed SFTSV infected cells on slides.…”

Section: Laboratory Diagnosismentioning

confidence: 99%

A highly pathogenic new bunyavirus emerged in China

2013

Emerging Microbes & Infections

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Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease that was discovered in China in 2010. The causative agent has been identified as a new member of the Phlebovirus genus in the family Bunyaviridae and has been designated severe fever with thrombocytopenia virus (SFTSV). SFTSV infection can be transmitted person-to-person, and the average case fatality rate is approximately 10% in humans. There is a high seroprevalence of SFTSV infection in a wide range of domesticated animals, including sheep, goats, cattle, pigs, dogs and chickens. Ticks are suspected to be the vector that transmits the virus to humans. Currently, the SFTS endemic area is expanding. Therefore, SFTSV infection is an increasingly important public health threat.

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Section: Laboratory Diagnosismentioning

confidence: 99%

A highly pathogenic new bunyavirus emerged in China

2013

Emerging Microbes & Infections

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“…Confirmation of SFTSV infection requires one of the following conditions: isolation of SFTSV from the patient's serum, detection of SFTSV RNA in the patient's serum by PCR, or detection of SFTS antibodies in the patient's serum. Several methods have been developed to amplify the viral RNA, including reverse transcription-PCR (RT-PCR) and RT-loop-mediated isothermal amplification (RT-LAMP) [29][30][31][32]. Amplification of the S segment is the most sensitive RT-PCR assay [29].…”

Section: Clinical Symptoms and Diagnosis Of Sftsmentioning

confidence: 99%

An emerging hemorrhagic fever in China caused by a novel bunyavirus SFTSV

Zhang

Liu

Zhao

et al. 2013

Sci. China Life Sci.

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Severe fever with thrombocytopenia syndrome (SFTS) is an emerging hemorrhagic fever in rural areas of China and is caused by a new bunyavirus, SFTSV, named after the disease. The transmission vectors and animal hosts of SFTSV are unclear. Ticks are the most likely transmission vectors and domestic animals, including goats, dogs, and cattle, are potential amplifying hosts of SFTSV. The clinical symptoms of SFTS are nonspecific, but major symptoms include fever, gastrointestinal symptoms, myalgia, dizziness, joint pain, chills, and regional lymphadenopathy. The most common abnormalities in laboratory test results are thrombocytopenia (95%), leukocytopenia (86%), and elevated levels of serum alanine aminotransferase, aspartate aminotransferase, creatine kinase, and lactate dehydrogenase. The fatality rate for SFTS is 12% on average, and the annual incidence of the disease is approximately five per 100000 of the rural population.

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“…Based on the nucleotide sequences of the Chinese SFTSV isolates, there have been some reports describing a reverse transcription-PCR (RT-PCR)-based method, a reverse transcription-loop-mediated isothermal amplification assay (RT-LAMP), reverse transcription-cross-priming amplification coupled (RT-CPA) with vertical flow (VF) visualization, and TaqMan-based quantitative real-time PCR for SFTSV detection (3,(6)(7)(8). However, since there are considerable sequence mismatches between the Chinese and Japanese lineages, especially in the reported TaqMan probe target regions, these primers/probes might not be suitable for use with the Japanese linage (4).…”

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confidence: 99%

Sensitive and Specific PCR Systems for Detection of Both Chinese and Japanese Severe Fever with Thrombocytopenia Syndrome Virus Strains and Prediction of Patient Survival Based on Viral Load

et al. 2014

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v Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease with a high case fatality risk and is caused by the SFTS virus (SFTSV).A retrospective study conducted after the first identification of an SFTS patient in Japan revealed that SFTS is endemic to the region, and the virus exists indigenously in Japan. Since the nucleotide sequence of Japanese SFTSV strains contains considerable differences compared with that of Chinese strains, there is an urgent need to establish a sensitive and specific method capable of detecting the Chinese and Japanese strains of SFTSV. A conventional one-step reverse transcription-PCR (RT-PCR) (cvPCR) method and a quantitative one-step RT-PCR (qPCR) method were developed to detect the SFTSV genome. Both cvPCR and qPCR detected a Chinese SFTSV strain. Forty-one of 108 Japanese patients suspected of having SFTS showed a positive reaction by cvPCR. The results from the samples of 108 Japanese patients determined by the qPCR method were in almost complete agreement with those determined by cvPCR. The analyses of the viral copy number level in the patient blood samples at the acute phase determined by qPCR in association with the patient outcome confirmed that the SFTSV RNA load in the blood of the nonsurviving patients was significantly higher than that of the surviving patients. Therefore, the cvPCR and qPCR methods developed in this study can provide a powerful means for diagnosing SFTS. In addition, the detection of the SFTSV genome level by qPCR in the blood of the patients at the acute phase may serve as an indicator to predict the outcome of SFTS.

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Development and evaluation of a reverse transcription loop-mediated isothermal amplification assay for rapid detection of a new SFTS bunyavirus

Cited by 32 publications

References 14 publications

A highly pathogenic new bunyavirus emerged in China

A highly pathogenic new bunyavirus emerged in China

An emerging hemorrhagic fever in China caused by a novel bunyavirus SFTSV

Sensitive and Specific PCR Systems for Detection of Both Chinese and Japanese Severe Fever with Thrombocytopenia Syndrome Virus Strains and Prediction of Patient Survival Based on Viral Load

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