scholarly journals Μελέτη του μηχανισμού παρασκευής in vitro ανασυνδυασμένων στελεχών πολιοϊών και εντεροϊών της ομάδας EV-C

2017 ◽  
Author(s):  
Τηλέμαχος Δημητρίου

Οι εντεροϊοί ανήκουν στην οικογένεια Picornaviridae. Το γένωμα τους είναι μονόκλωνο RNA θετικής πολικότητας μήκους περίπου 7.500bp και περιβάλλεται από ένα εικοσαεδρικό πρωτεϊνικό καψίδιο. Οι εντεροϊοί που προσβάλλουν τον άνθρωπο ταξινομούνται σε τέσσερις ομάδες: EV-A, EV-B, EV-C και EV-D. Οι πολιοϊοί, το σημαντικότερο μέλος της ομάδας C, διακρίνονται σε τρεις ορότυπους (PV1, PV2, PV3) και είναι οι αιτιολογικοί παράγοντες της παραλυτικής πολιομυελίτιδας. Από το 1960 χρησιμοποιούνται δύο εμβόλια για την εξάλειψη της ασθένειας, αρχικά το IPV (inactivated polio vaccine) και κατόπιν το πιο αποτελεσματικό OPV (oral polio vaccine). Ωστόσο, το OPV εμφάνισε το μειονέκτημα της εμβολιοσυνδεόμενης παραλυτικής πολιομυελίτιδας (VAPP: Vaccine-associated paralytic poliomyelitis) και της κυκλοφορίας των εμβολιοπροερχόμενων πολιοϊών (VDPVs: Vaccine Derived Polioviruses) μέσω της συσσώρευσης μεταλλάξεων ή και ανασυνδυασμών στο γένωμα των εξασθενημένων εμβολιακών στελεχών Sabin.Στόχος της παρούσας διατριβής ήταν ο σχεδιασμός και η ανάπτυξη μεθόδων με σκοπό τη μελέτη του μηχανισμού παρασκευής in vitro ανασυνδυασμένων στελεχών πολιοϊών και εντεροϊών της ομάδας EV-C. Στο πρώτο μέρος της διατριβής σχεδιάστηκε και αναπτύχθηκε μια Multiplex-PCR για την ανίχνευση και ταυτοποίηση των εντεροϊών. Τα αποτελέσματα της τεχνικής αυτής σε πρότυπα αλλά και κλινικά στελέχη ανέδειξαν την τεχνική αυτή ως ένα χρήσιμο εργαλείο για την γρήγορη και ακριβή ανίχνευση και ταυτοποίηση των εντεροϊών.Στο δεύτερο μέρος της διατριβής, σχεδιάστηκε και αναπτύχθηκε μια τεχνική αλληλούχησης ολόκληρου του γονιδιώματος των εντεροϊών χρησιμοποιώντας μόνο τέσσερις PCR αντιδράσεις. Η δυνατότητα αυτή παρέχεται μέσω της χρήσης ενός ειδικού εκκινητή (DOP), μέσω του οποίου πραγματοποιήθηκε αρχικά μια προενίσχυση ολόκληρου του γονιδιώματος. Στο τρίτο μέρος της διατριβής σχεδιάστηκε και εφαρμόστηκε σε κλινικά δείγματα μια Multiplex-PCR για την ανίχνευση ανασυνδυασμών από τη VP1 έως και τη 3D γενωμική περιοχή εμβολιοσυνδεόμενων πολιοϊών. Τα αποτελέσματα της Multiplex-PCR απέδειξαν την ικανότητα της μεθόδου να ανιχνεύει και να ταυτοποιεί σπάνιους αλλά και κύριους τύπους ανασυνδυασμού με τη χρήση μόνο τεσσάρων Multiplex-PCR αντιδράσεων.Στο τέταρτο μέρος της διατριβής σχεδιάστηκε μια ειδική Stem-Loop RT-PCR μέθοδος για την ανίχνευση της αντιγραφικής ενεργότητας των εντεροϊών μέσω ανίχνευσης του αρνητικής πολικότητας RNA κλώνου. Η μέθοδος αυτή εφαρμόστηκε αρχικά στο πρότυπο στέλεχος Sabin 1, όπου ανιχνεύτηκε η αντιγραφική ενεργότητα του στελέχους σε υψηλό αλλά και χαμηλό ιικό τίτλο, αρκετά νωρίτερα από την εμφάνιση της χαρακτηριστικής εικόνας CPE, των ενεργών εντεροϊών σε κυτταροκαλλιέργεια. Επιπλέον, η μέθοδος αυτή χρησιμοποιήθηκε για την διάκριση μεταξύ αντιγραφικά ενεργών και ανενεργών πρότυπων στελεχών CAV που δεν εμφάνιζαν CPE σε κυτταροκαλλιέργειες. Στο τελευταίο μέρος της παρούσας διατριβής πραγματοποιήθηκε η μελέτη των ανασυνδυασμών που προέκυψαν έπειτα από ταυτόχρονη μόλυνση κυττάρων Rd με πρότυπο στέλεχος Sabin 1 και CAV13. Παράλληλα, με τη χρήση ειδικών προγραμμάτων βιοπληροφορικής σχεδιάστηκαν πιθανά μοντέλα της δευτεροταγούς δομής των RNA μορίων στις θέσεις ανασυνδυασμού. Τα αποτελέσματα της μελέτης αυτής επαλήθευσαν την αυξημένη συχνότητα εμφάνισης ετεροτυπικών ανασυνδυασμών στην 2Α ή 2Β γενωμική περιοχή και εμφάνισαν μια ένδειξη που επιβεβαιώνει τη σύνδεση μεταξύ ανασυνδυασμού και δευτεροταγούς δομής του RNA μορίου.

2021 ◽  
Vol 308 ◽  
pp. 02018
Author(s):  
Yushuo Chen ◽  
Tianrui Yue ◽  
Zixiao Zhang

Poliomyelitis is an exclusively human disease that mainly affects children. Clinical features of poliomyelitis can be varied, from mild illness to the most severe paralysis, and the factor why poliomyelitis has different performances in individuals has been proved strongly correlated with membrane protein CD155. The nervous system shows a special protecting phenomenon against the invasion of poliovirus, and the mechanism is not very clear at present. Vaccines are the main means of preventing and controlling polio, and many different vaccines have been invented in the process of fighting polio. Inactivated polio vaccine (IPV) and oral polio vaccine (OPV) are the two main vaccines. IPV is known for its safety while OPV is widely used in developing countries because of its relatively low cost. This usage also leads to some side effects: vaccine-associated paralytic polio (VAPP) and vaccine-derived poliovirus (VDPV). Now, for polio eradication, the elimination of these two diseases has become particularly important. Thus, a new type of vaccine was created: sequential IPV-OPV with the safety of IPV and the low cost of OPV. This paper will talk about the different polio vaccines and their effects. An enormous difference between people who have gotten the vaccine and people who have not got the vaccine. Comparing the two kinds of people, people who get normal poliovirus, and people who get poliovirus after taking a vaccine, known as VAPP (vaccine-associated paralytic poliomyelitis), the former cannot get full recovery whole life and the latter has a very low possibility. In conclusion, people should take vaccines if it is affordable for them.


2000 ◽  
Vol 124 (1) ◽  
pp. 113-120 ◽  
Author(s):  
D. RIDGWAY

Beginning in January 1997, American immunization policy allowed parents and physicians to elect one of three approved infant vaccination strategies for preventing poliomyelitis. Although the three strategies likely have different outcomes with respect to prevention of paralytic poliomyelitis, the extreme rarity of the disease in the USA prevents any controlled comparison. In this paper, a formal inferential logic, originally described by Donald Rubin, is applied to the vaccination problem. Assumptions and indirect evidence are used to overcome the inability to observe the same subjects under varying conditions to allow the inference of causality from non-randomized observations. Using available epidemiologic information and explicit assumptions, it is possible to project the risk of paralytic polio for infants immunized with oral polio vaccine (1·3 cases per million vaccinees), inactivated polio vaccine (0·54 cases per million vaccinees), or a sequential schedule (0·54–0·92 cases per million vaccinees).


2018 ◽  
Vol 67 (suppl_1) ◽  
pp. S57-S65 ◽  
Author(s):  
James T Gaensbauer ◽  
Chris Gast ◽  
Ananda S Bandyopadhyay ◽  
Miguel O’Ryan ◽  
Xavier Saez-Llorens ◽  
...  

2020 ◽  
Vol 35 (Supplement_1) ◽  
pp. i30-i37
Author(s):  
Mirembe Rachel Faith ◽  
Babirye Juliet ◽  
Nathan Tumuhamye ◽  
Tumwebaze Mathias ◽  
Emma Sacks

Abstract Uganda officially introduced the inactivated polio vaccine (IPV) in May 2016 as part of the polio eradication strategy and integrated it into its routine immunization programme in addition to the oral polio vaccine. The current coverage stands at 60% as of July 2017. We therefore aimed to determine factors associated with the uptake of IPV among children in Kalungu District so as to inform the implementation of the vaccine policy. A community-based cross-sectional study was conducted among caregivers of 406 eligible children aged 12–23 months through multi-stage systematic sampling and a standardized semi-structured questionnaire. Nine key informant interviews were conducted through purposive selection of health care providers and members of Village Health Teams (VHTs) based on their expertize. Modified Poisson regression and thematic content analysis were used to determine factors significant to IPV uptake among children. 71% of sampled children aged 12–23 months had received IPV in Kalungu District. The survey found that being encouraged by health workers and VHTs was significant to children’s uptake of IPV (Adjusted PR 1.24, 95% CI; 1.22–3.47). Distance to the immunization point (Adjusted PR 0.32,95% CI; 0.16–0.62) and caregiver’s education level (Adjusted PR 1.16,95% CI; 1.05–2.22) were also associated with IPV uptake. Qualitative findings from health workers and VHT members further confirmed the perception that distance to the immunization post was important, and VHTs also stated that being encouraged by health workers was critical to IPV uptake. The current prevalence of IPV uptake among children aged 12–23 months in Kalungu is 71%, higher than the last reported national coverage (60%), though still below the recommended national coverage of 95%. Efforts should be focused on sensitization of caregivers through health workers and VHTs. Immunization outreach should be strengthened so as to bring services closer to patients.


2007 ◽  
Vol 136 (2) ◽  
pp. 180-183 ◽  
Author(s):  
H. E. GARY ◽  
B. SMITH ◽  
J. JENKS ◽  
J. RUIZ ◽  
W. SESSIONS ◽  
...  

SUMMARYWhile oral polio vaccine (OPV) has been shown to be safe and effective, it has been observed that it can circulate within a susceptible population and revert to a virulent form. Inactivated polio vaccine (IPV) confers protection from paralytic disease, but provides limited protection against infection. It is possible, then, that an IPV-immunized population, when exposed to OPV, could sustain undetected circulation of vaccine-derived poliovirus. This study examines the possibility of polio vaccine virus circulating within the United States (highly IPV-immunized) population that borders Mexico (OPV-immunized). A total of 653 stool and 20 sewage samples collected on the US side of the border were tested for the presence of poliovirus. All samples were found to be negative. These results suggest that the risk of circulating vaccine-derived poliovirus is low in fully immunized IPV-using populations in developed countries that border OPV-using populations.


2015 ◽  
Vol 49 (0) ◽  
Author(s):  
Ana Marli Christovam Sartori ◽  
Margarete Paganotti Vicentine ◽  
Lígia Castelloni Figueiredo Gryninger ◽  
Patricia Coelho de Soárez ◽  
Hillegonda Maria Dutilh Novaes

OBJECTIVE To analyze the costs of vaccination regimens for introducing inactivated polio vaccine in routine immunization in Brazil.METHODS A cost analysis was conducted for vaccines in five vaccination regimens, including inactivated polio vaccine, compared with the oral polio vaccine-only regimen. The costs of the vaccines were estimated for routine use and for the “National Immunization Days”, during when the oral polio vaccine is administered to children aged less than five years, independent of their vaccine status, and the strategic stock of inactivated polio vaccine. The presented estimated costs are of 2011.RESULTS The annual costs of the oral vaccine-only program (routine and two National Immunization Days) were estimated at US$19,873,170. The incremental costs of inclusion of the inactivated vaccine depended on the number of vaccine doses, presentation of the vaccine (bottles with single dose or ten doses), and number of “National Immunization Days” carried out. The cost of the regimen adopted with two doses of inactivated vaccine followed by three doses of oral vaccine and one “National Immunization Day” was estimated at US$29,653,539. The concomitant replacement of the DTPw/Hib and HepB vaccines with the pentavalent vaccine enabled the introduction of the inactivated polio without increasing the number of injections or number of visits needed to complete the vaccination.CONCLUSIONS The introduction of the inactivated vaccine increased the annual costs of the polio vaccines by 49.2% compared with the oral vaccine-only regimen. This increase represented 1.13% of the expenditure of the National Immunization Program on the purchase of vaccines in 2011.


2021 ◽  
Vol 1 (12) ◽  
Author(s):  
Wahyu Fitrah Darwanto Nugroho ◽  
Indra Kusuma ◽  
Siti Nur Riani

Latar Belakang : Vaksin merupakan suspensi mikroorganisme yang dilemahkan atau dimatikan, atau protein antikgenik dari berbagai organisme tadi yang diberikan untuk mencegah, meringankan, atau mengobati penyakit-penyakit menular. Vaksin pertama kali tercatat pada tahun 1769, yang dipublikasikan oleh Edward Jenner, yaitu specimen yang berasal dari lesi lengan seseorang yang terinfeksi Cowpox. Human Diploid Cells (HDC) merupakan salah satu sel yang digunakan untuk mengkultur virus yang akan dijadikan vaksin. HDC yang berasal dari aborsi manusia ini banyak digunakan untuk mengkultur virus Polio IPV dan OPV, Rabies, Rubella, Measles, Varicella-Zooster, dan Hepatitis A. Tujuan : Vaksin polio merupakan vaksin yang diwajibkan pada anak yang dijadwalkan dari Ikatan Dokter Anak Indonesia (IDAI) yang dibagi menjadi dua jenis, IPV (Inactivated Polio Vaccine) dan OPV (Oral Polio Vaccine). Metode : Jenis Penelitian yang digunakan adalah deskriptif dengan pendekatan cross sectional menggunakan kuesioner. Populasi yang digunakan adalah mahasisa Fakultas Kedokteran Universitas YARSI tahun pertama dan tahun ketiga yang memenuhi syarat. Cara pemilihan sampel dengan simple random sampling. Hasil : Penelitian yang dilaksanakan selama 3 hari dengan menggunakan kuesioner, dari 100 responden didapatkan persentase jumlah kuesioner Pengetahuan mengenai Human Diploid Cell berdasarkan Tingkat Pendidikan didapatkan pengetahuan baik sebanyak 5% pada tahun ketiga dan 7% pada tahun pertama. Pengetahuan cukup sebanyak 23% pada tingkat ketiga dan 28% pada tahun pertama. Pengetahuan kurang sebanyak 9% pada tingkat ketiga dan 28% pada tahun pertama. Persentase jumlah kuesioner Pengetahuan mengenai Polio berdasarkan Tingkat Pendidikan didapatkan pengetahuan baik sebanyak 15% pada tahun ketiga dan 19% pada tahun pertama. Pengetahuan cukup sebanyak 18% pada tingkat ketiga dan 31% pada tahun pertama. Pengetahuan kurang sebanyak 4% pada tingkat ketiga dan 13% pada tahun pertama. Kesimpulan : Tidak terdapat hubungan antara tingkat pendidikan dengan pengetahuan mengenai Human Diploid Cell dalam vaksin Polio. Dalam pandangan Islam, penggunaan vaksin Polio hukumnya mubah karena prinsip Dharuriyat bertujuan untuk mempertahankan nyawa atau Hifdz an-nafs anak dari ancaman penyakit.


2020 ◽  
Vol 41 (4) ◽  
pp. 196
Author(s):  
Margaret M Peel

Epidemics of paralytic poliomyelitis (polio) first emerged in the late 19th and early 20th centuries in the United States and the Scandinavian countries. They continued through the first half of the 20th century becoming global. A major epidemic occurred in Australia in 1951 but significant outbreaks were reported from the late 1930s to 1954. The poliovirus is an enterovirus that is usually transmitted by the faecal–oral route but only one in about 150 infections results in paralysis when the central nervous system is invaded. The Salk inactivated polio vaccine (IPV) became available in Australia in 1956 and the Sabin live attenuated oral polio vaccine (OPV) was introduced in 1966. After decades of stability, many survivors of the earlier epidemics experience late-onset sequelae including post-polio syndrome. The World Health Organization launched the global polio eradication initiative (GPEI) in 1988 based on the easily administered OPV. The GPEI has resulted in a dramatic decrease in cases of wild polio so that only Pakistan and Afghanistan report such cases in 2020. However, a major challenge to eradication is the reversion of OPV to neurovirulent mutants resulting in circulating vaccine-derived poliovirus (cVDPV). A novel, genetically stabilised OPV has been developed recently to stop the emergence and spread of cVDPV and OPV is being replaced by IPV in immunisation programs worldwide. Eradication of poliomyelitis is near to achievement and the expectation is that poliomyelitis will join smallpox as dreaded epidemic diseases of the past that will be consigned to history.


2020 ◽  
Vol 41 (4) ◽  
pp. 223
Author(s):  
Margaret M Peel

Epidemics of paralytic poliomyelitis (polio) first emerged in the late 19th and early 20th centuries in the United States and the Scandinavian countries. They continued through the first half of the 20th century becoming global. A major epidemic occurred in Australia in 1951 but significant outbreaks were reported from the late 1930s to 1954. The poliovirus is an enterovirus that is usually transmitted by the faecal–oral route but only one in about 150 infections results in paralysis when the central nervous system is invaded. The Salk inactivated polio vaccine (IPV) became available in Australia in 1956 and the Sabin live attenuated oral polio vaccine (OPV) was introduced in 1966. After decades of stability, many survivors of the earlier epidemics experience late-onset sequelae including post-polio syndrome. The World Health Organization launched the global polio eradication initiative (GPEI) in 1988 based on the easily administered OPV. The GPEI has resulted in a dramatic decrease in cases of wild polio so that only Pakistan and Afghanistan report such cases in 2020. However, a major challenge to eradication is the reversion of OPV to neurovirulent mutants resulting in circulating vaccine-derived poliovirus (cVDPV). A novel, genetically stabilised OPV has been developed recently to stop the emergence and spread of cVDPV and OPV is being replaced by IPV in immunisation programs worldwide. Eradication of poliomyelitis is near to achievement and the expectation is that poliomyelitis will join smallpox as dreaded epidemic diseases of the past that will be consigned to history.


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