Investigation of Adenovirus Isolation Frequency from the Stool Samples of Patients Suspected with Acute Flaccid Paralysis

ABSTRACT We analyzed viral nucleic acids in stool samples collected from 35 South Asian children with nonpolio acute flaccid paralysis (AFP). Sequence-independent reverse transcription and PCR amplification of capsid-protected, nuclease-resistant viral nucleic acids were followed by DNA sequencing and sequence similarity searches. Limited Sanger sequencing (35 to 240 subclones per sample) identified an average of 1.4 distinct eukaryotic viruses per sample, while pyrosequencing yielded 2.6 viruses per sample. In addition to bacteriophage and plant viruses, we detected known enteric viruses, including rotavirus, adenovirus, picobirnavirus, and human enterovirus species A (HEV-A) to HEV-C, as well as numerous other members of the Picornaviridae family, including parechovirus, Aichi virus, rhinovirus, and human cardiovirus. The viruses with the most divergent sequences relative to those of previously reported viruses included members of a novel Picornaviridae genus and four new viral species (members of the Dicistroviridae, Nodaviridae, and Circoviridae families and the Bocavirus genus). Samples from six healthy contacts of AFP patients were similarly analyzed and also contained numerous viruses, particularly HEV-C, including a potentially novel Enterovirus genotype. Determining the prevalences and pathogenicities of the novel genotypes, species, genera, and potential new viral families identified in this study in different demographic groups will require further studies with different demographic and patient groups, now facilitated by knowledge of these viral genomes.

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Preponderance of Enterovirus Species C in RD-L20B cell culture negative stool samples from children diagnosed with Acute Flaccid Paralysis in Nigeria

10.1101/116004 ◽

2017 ◽

Author(s):

J.A. Adeniji ◽

A.O. Oragwa ◽

U.E. George ◽

U.I. Ibok ◽

T.O.C. Faleye ◽

...

Keyword(s):

Cell Culture ◽

Acute Flaccid Paralysis ◽

Direct Detection ◽

Rna Extraction ◽

Clinical Specimen ◽

Flaccid Paralysis ◽

Surveillance Program ◽

Two Samples ◽

Afp Surveillance ◽

Stool Samples

AbstractsRecently, a reverse transcriptase seminested polymerase chain reaction (RT-snPCR) assay was recommended by the WHO for direct detection of enteroviruses from clinical specimen. In this study, we use the assay and its modification to screen acute flaccid paralysis (AFP) samples previously confirmed negative for enteroviruses by the RD-L20B algorithm.Thirty paired stool suspensions collected in 2015 as part of the national AFP surveillance program in different states of Nigeria were analyzed in this study. The samples were previously confirmed negative for enteroviruses by the polio laboratory in accordance with the WHO recommended RD-L20B cell culture based algorithm. Two samples previously confirmed to contain enteroviruses were included as positive controls. All samples were subjected to RNA extraction, and the RT-snPCR assay and its modifications. All amplicons were sequenced and enteroviruses identified using the enterovirus genotyping tool.Overall, amplicons were recovered from the two controls and 50% (15/30) of samples screened. Fourteen were successfully typed of which, 7.1% (1/14), 21.4% (3/14), 64.3% (9/14) and 7.1% (1/14) were EV-A, EV-B, EV-C and a mixture of EV-B and C (EV-C99 and E25), respectively. The two controls were identified as EV-C99 and CV-A1, both EV-Cs. The PV-2 detected had VP1 ILE143. Hence, a vaccine strain.The results of this study showed that about 50% of enterovirus infections (including some Sabin PV2s) are being missed by the RD-L20B cell culture based algorithm. This highlights the value of the RT-snPCR assay and its modifications. The circulation and preponderance of EV-Cs in Nigeria was also confirmed.

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Faculty Opinions recommendation of Metagenomic analyses of viruses in stool samples from children with acute flaccid paralysis.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1159145.619446 ◽

2009 ◽

Author(s):

Lynn Enquist

Keyword(s):

Acute Flaccid Paralysis ◽

Flaccid Paralysis ◽

Stool Samples

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Development of a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) system for a highly sensitive detection of enterovirus in the stool samples of acute flaccid paralysis cases

BMC Infectious Diseases ◽

10.1186/1471-2334-9-208 ◽

2009 ◽

Vol 9 (1) ◽

Cited By ~ 17

Author(s):

Minetaro Arita ◽

Hua Ling ◽

Dongmei Yan ◽

Yorihiro Nishimura ◽

Hiromu Yoshida ◽

...

Keyword(s):

Reverse Transcription ◽

Acute Flaccid Paralysis ◽

Isothermal Amplification ◽

Sensitive Detection ◽

Flaccid Paralysis ◽

Loop Mediated Isothermal Amplification ◽

Highly Sensitive ◽

Stool Samples ◽

Lamp System ◽

Highly Sensitive Detection

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Preponderance of enterovirus C in RD-L20B-cell-culture-negative stool samples from children diagnosed with acute flaccid paralysis in Nigeria

Archives of Virology ◽

10.1007/s00705-017-3466-2 ◽

2017 ◽

Vol 162 (10) ◽

pp. 3089-3101 ◽

Cited By ~ 12

Author(s):

J. A. Adeniji ◽

A. O. Oragwa ◽

U. E. George ◽

U. I. Ibok ◽

T. O. C. Faleye ◽

...

Keyword(s):

Cell Culture ◽

Acute Flaccid Paralysis ◽

Flaccid Paralysis ◽

Stool Samples ◽

Culture Negative

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Cases of Acute Flaccid Paralysis Associated with Coxsackievirus A2: Findings of a 20-Year Surveillance in the Russian Federation

Microorganisms ◽

10.3390/microorganisms10010112 ◽

2022 ◽

Vol 10 (1) ◽

pp. 112

Author(s):

Olga E. Ivanova ◽

Armen K. Shakaryan ◽

Nadezhda S. Morozova ◽

Yulia A. Vakulenko ◽

Tatyana P. Eremeeva ◽

...

Keyword(s):

Acute Flaccid Paralysis ◽

Laboratory Data ◽

Foot And Mouth Disease ◽

Polio Eradication ◽

Vaccination Schedule ◽

Flaccid Paralysis ◽

Global Polio Eradication Initiative ◽

Afp Surveillance ◽

Stool Samples ◽

The Russian Federation

Surveillance for acute flaccid paralysis syndrome (AFP) in children under 15 is the backbone of the Global Polio Eradication Initiative. Laboratory examination of stool samples from AFP cases allows the detection of, along with polioviruses, a variety of non-polio enteroviruses (NPEV). The etiological significance of these viruses in the occurrence of AFP cases has been definitively established only for enteroviruses A71 and D68. Enterovirus Coxsackie A2 (CVA2) is most often associated with vesicular pharyngitis and hand, foot and mouth disease. Among 7280 AFP cases registered in Russia over 20 years (2001–2020), CVA2 was isolated only from five cases. However, these included three children aged 3 to 4 years, without overt immune deficiency, immunized with 4–5 doses of poliovirus vaccine in accordance with the National Vaccination Schedule. The disease resulted in persistent residual paralysis. Clinical and laboratory data corresponded to poliomyelitis developing during poliovirus infection. These findings are compatible with CVA2 being the cause of AFP. Molecular analysis of CVA2 from these patients and a number of AFP cases in other countries did not reveal association with a specific phylogenetic group, suggesting that virus genetics is unlikely to explain the pathogenic profile. The overall results highlight the value of AFP surveillance not just for polio control but for studies of uncommon AFP agents.

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Abundance of Enterovirus C in RD-L20B cell culture negative stool samples from Acute Flaccid Paralysis cases in Nigeria is geographically defined

10.1101/222935 ◽

2017 ◽

Author(s):

E Donbraye ◽

O.I. Olasunkanmi ◽

B.A. Opabode ◽

T.R. Ishola ◽

T.O.C. Faleye ◽

...

Keyword(s):

Cell Culture ◽

Phylogenetic Analysis ◽

Acute Flaccid Paralysis ◽

Rna Extraction ◽

Geographical Region ◽

Flaccid Paralysis ◽

Cdna Synthesis ◽

Faecal Samples ◽

Stool Samples ◽

Culture Negative

ABSTRACTWe recently showed that Enteroviruses (EVs); majorly species Cs (EV-Cs) were present in about 46.7% of faecal samples from children <15 years old diagnosed with Acute Flaccid Paralysis (AFP) in Nigeria but declared to be EV free by the RD-L20B cell culture based algorithm. In this study, we investigated whether this observed preponderance of EVs (and EV-Cs) in such samples vary by geographical region.In all, 108 samples (i.e. 54 paired stool suspensions from 54 AFP cases) previously confirmed negative for EVs by the WHO recommended RD-L20B cell culture based algorithm were analyzed in this study. The 108 samples were made into 54 pools (27 each from Northwest [NW] and Southsouth [SS] Nigeria). All samples were subjected to RNA extraction, cDNA synthesis and the WHO recommended seminestedPCR (snPCR) assay and its modifications. All amplicons were sequenced, and enteroviruses identified using the enterovirus genotyping tool and phylogenetic analysis.Altogether, EVs were detected in 16 (29.63%) of the 54 samples screened but successfully identified in 14 (25.93%): 10 from NW- and 4 from SS-Nigeria. Precisely, one (7.14%), two (14.29%) and 11 (78.57%) of the strains detected were EV-A, EV-B and EV-C respectively. The 10 strains from NW-Nigeria are 7 EV types and include CV-A10, E29, CV-A13, CV-A17, CV-A19, CV-A24 and EV-C99. The four EV types recovered from SS-Nigeria include E31, CV-A1, EV-C99 and EV-C116. EV-C99 is the only EV type that was detected in both NW- and SS-Nigeria.The results of this study showed that the preponderance of EVs and consequently EV-Cs in AFP samples declared to be EV free by the RD-L20B cell culture based algorithm vary by geographical region in Nigeria. It further confirmed the EV-B bias of the RD-L20B cell culture based algorithm.

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Molecular Characterization of Enteroviruses Isolated from Acute Flaccid Paralysis Cases in Poland, 1999–2014

Polish Journal of Microbiology ◽

10.5604/17331331.1227670 ◽

2016 ◽

Vol 65 (4) ◽

pp. 443-450 ◽

Cited By ~ 3

Author(s):

Magdalena Wieczorek ◽

Arleta Krzysztoszek

Keyword(s):

Acute Flaccid Paralysis ◽

Wide Distribution ◽

Flaccid Paralysis ◽

Viral Pathogens ◽

The Past ◽

Different Types ◽

Afp Surveillance ◽

Stool Samples ◽

Healthy Contact

Enteroviruses (EVs) are among viral pathogens that can cause acute flaccid paralysis (AFP). This study represents an overview of EVs isolated through AFP surveillance in Poland between 1999 and 2014. The presence of enteroviruses was studied in stool samples that were collected from 747 AFP cases and their asymptomatic contacts. Fifty five (6.12%) cases of AFP were associated with enterovirus isolation. Out of the 55 positive cases, 40 were associated with detection of enterovirus in patient, and 15 with detection of EV in healthy contact, without positive detection in paralytic patient. Polioviruses were isolated from 35 AFP cases. The results of this study showed that about 43.6% of positive AFP cases were found in association with the isolation of non-polio enteroviruses (NPEV). A total of 12 different types of the species B were detected (CVA9, CVB1, CVB3, CVB4, CVB5, E3, E4, E9, E11, E13, E30), and one additional isolate represented the species enterovirus A (EV71). Among the 12 serotypes of species B, CVB3 and CVB5 were more frequently detected than others, representing 40% of the characterized isolates, followed by CVB4 (16%), E4 (8%), and E11(8%). Phylogenetic analysis revealed that strains from Poland had the closest genetic relationship with isolates previously identified in Europe (France, Finland, Denmark, Moldova) but also in other parts of the world (Tunisia, China, USA), suggesting wide distribution of these lineages. The paper provides information about NPEV circulation in Poland in the past 16 years, about its association with the AFP and it indicates the need for monitoring NPEV circulation even after the eradication of poliomyelitis.

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