Detection and Typing of Human Papilloma Virus by Multiplex PCR with Type-Specific Primers

Pastrana, Francisco Romero

doi:10.5402/2012/186915

Cited by 27 publications

(29 citation statements)

References 26 publications

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“…In this study less than 50% of the women were infected with one or more HPV genotypes, which were heterogeneously distributed irrespective of their oncogenic risk. Our results are comparable with those from other studies with Mexican women [11] [18] [19]. In contrast, a meta-analysis about worldwide HPV detection among women with normal cytological findings, revealed an estimated frequency for Central America (including Mexico) of around 20%, while estimated frequencies for worldwide and developing region were 7.2% and 14.3%, respectively [20].…”

Section: Discussionsupporting

confidence: 57%

“…On the contrary, when groups of Mexican women presenting LSIL, HSIL or cervical cancer were analyzed, frequencies of HPV infection increased from 42% to 95%, irrespective of the lesion type [19], although individual high-risk genotype frequencies were variable between studies. This confirms that there is a high correlation between presence of oncogenic HPV genotypes and different cervical lesions [2] [4].…”

Section: Discussionmentioning

confidence: 99%

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Relevant Frequency of Multiple Infections with High- and Low-Risk HPV Genotypes among Mexican Women Attending a Tertiary Care Hospital

Flores-Medina¹,

García-Romero²,

Soriano-Becerril³

et al. 2016

OJOG

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Aims: To assess the frequency and the main HPV genotypes circulating among a group of women attending at a third level Hospital in Mexico City. Methods: A cross-sectional and descriptive study was performed in a group of 143 female outpatients of the Gynecology and Obstetrics Service at the National Institute of Perinatology of Mexico. Cervical swabs were taken from participants and subjected to simultaneous detection/genotyping of HPV by Linear Array Genotyping Test (Roche Molecular Systems). Mann-Whitney U, median and/or Square Chi tests were used to compare socio-demographical features between HPV-infected and uninfected women. Results: A total of 66 women (46.2%) had HPV infection. Overall, 112 genotypes were detected either as single infections (45.5%) or multiple genotype infections (54.5%). The cumulated frequency of multiple infections with high-/low-and high-/high-risk HPV genotypes was 63.9 %. The most frequent high-risk genotypes were HPV52 HPV58 and HPV51, whereas the most frequent low-risk genotypes were HPV6, HPV53 and HPV84. Infected women were significantly younger and have less stable partner relationships than uninfected women (p < 0.05). Conclusion: A relevant frequency of mixed infections with high-and low-risk HPV genotypes, other than those consi-* Corresponding author. S. Flores-Medina et al.425 dered most prevalent worldwide, was observed. Most circulating high-risk genotypes among the women of this study are not covered by commercial vaccine formulations.

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Section: Discussionsupporting

confidence: 57%

Section: Discussionmentioning

confidence: 99%

Relevant Frequency of Multiple Infections with High- and Low-Risk HPV Genotypes among Mexican Women Attending a Tertiary Care Hospital

Flores-Medina¹,

García-Romero²,

Soriano-Becerril³

et al. 2016

OJOG

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“…Our study shows that to minimize the damage to insect farming caused by pathogenic microorganisms, multiplex PCR-based detection and capillary electrophoresisbased quantification may be an effective and rapid approach to identify and prevent spreading of insect disease and massive insect death. Also, rapid and sensitive diagnosis of insect disease is necessary to promote further industrialization of insect farming and to obtain insect-derived biologically active substances.Compared to single PCR, multiplex PCR demonstrates advantages as a fast and cost-effective method for simultaneous detection of several insect pathogens; however, multiplex PCR has limitations when quantitative evaluation of insect co-infection is required or the contribution of each pathogen to disease progress should be assessed (De Smet et al, 2012; Khare et al, 2014;Romero-Pastrana, 2012; Sguazza et al, 2013b). In such cases, capillary electrophoresis can be used as a sensitive method to quantitatively analyze and compare the presence of each target pathogen gene (Barakat et al, 2014;Chung et al, 2012;Shin et al, 2014;Shin et al, 2010).…”

mentioning

confidence: 99%

“…Compared to single PCR, multiplex PCR demonstrates advantages as a fast and cost-effective method for simultaneous detection of several insect pathogens; however, multiplex PCR has limitations when quantitative evaluation of insect co-infection is required or the contribution of each pathogen to disease progress should be assessed (De Smet et al, 2012; Khare et al, 2014;Romero-Pastrana, 2012; Sguazza et al, 2013b). In such cases, capillary electrophoresis can be used as a sensitive method to quantitatively analyze and compare the presence of each target pathogen gene (Barakat et al, 2014;Chung et al, 2012;Shin et al, 2014;Shin et al, 2010).…”

mentioning

confidence: 99%

Simultaneous detection of fungal, bacterial, and viral pathogens in insects by multiplex PCR and capillary electrophoresis

Kwak¹,

Nam²,

Choi³

et al. 2015

International Journal of Industrial Entomology

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Beetles Protaetia brevitarsis seulensis Kolbe (Coleoptera: Cetoniidae) and Allomyrina dichotoma Linn. (Coleoptera: Scarabaeidae) are widely used in traditional medicine, and the number of insect-rearing farms is increasing in South Korea. The purpose of this study was to establish a multiplex PCR-based assay for rapid simultaneous detection of multiple pathogens causing insect diseases. Six insect parasites such as fungi Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Cordycipitaceae) and Metarhizium anisopliae (Metschn.) Sorokin (Hypocreales: Clavicipitaceae), bacteria Bacillus thuringiensis Berliner (Bacillales: Bacillaceae), Pseudomonas aeruginosa Migula (Pseudomonadales: Pseudomonadaceae), and Serratia marcescens Bizio (Enterobacteriales: Enterobacteriaceae), and Oryctes rhinoceros nudivirus were chosen based on the severity and incidence rate of insect diseases in South Korea. Pathogen-specific primers were designed and successfully applied for simultaneous detection of multiple infectious agents in farm-bred insects P. b. seulensis and A. dichotoma using multiplex PCR and high resolution capillary electrophoresis. Our results indicate that multiplex PCR is an effective and time-saving method for simultaneous detection of multiple infections in insects, and the QIAxcel capillary electrophoresis system is useful for quantitative evaluation of the individual impact of each infectious agent on the severity of insect disease. The approach designed in this study can be utilized for rapid and accurate diagnostics of infection in insect farms.© 2015 The Korean Society of Sericultural Sciences Int. J. Indust. Entomol. 30(2), 64-74 (2015) IntroductionFollowing a global trend of using natural products from insects, the Korean government has been encouraging the development of insect resources for environmental and dietary purposes, traditional medicine, and as pets (Kim et al., 2013 Vol. 30, No. (1), pp. 64-74 (2015) Pseudomonas aeruginosa is reported as a virulent bacterial pathogen not only for insects but also for mammals (Jander et al., 2000), where it causes infection of the respiratory tract, especially in cystic fibrosis patients (Spiker et al., 2004). Similar to Pseudomonas aeruginosa, Serratia marcescens is known as an opportunistic bacterium (El-Aasar et al., 2013); it is also used to control apple maggot fly population and as a source of insecticidal agents such as toxin luxA (Hejazi and Falkiner, 1997; Lauzon et al., 2003). Oryctes rhinoceros virus Rhabdionvirus oryctes Hüger was first discovered in oil palms in Malaysia 40 years ago and is currently used for the control of rhinoceros beetle population in Malaysia (Huger, 2005;Ramle et al., 2005).We have identified this virus in an A. dichotoma-breeding farm in 2014 in Korea (Lee, 2015).Extensive large-scale insect breeding has resulted in frequent occurrence of insect infection due to multiple pathogens. Here, we developed a multiplex PCR-based approach that allowed simultaneous detection of six insect pathogens using specific primer se...

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“…Cervical cancer is highly prevalent in the developing and poor countries and it is said to include 80% of the total cancers women (2). This cancer is known to be the second prevalent cancer among Iranian women (3).…”

mentioning

confidence: 99%

Qualitative-based Multiplex PCR: Diagnosis of Papilloma virus Types 16 & 18 in Samples Taken from Patients with Malignant Lesions

Shafaati¹,

Akhavan²,

Yazdani³

et al. 2015

Iran J Virol

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Background and Aims: The human papilloma virus was introduced as the major etiological agent in outbreak of cervical cancer in 1970. Since it is very difficult to recognize these viruses and their types using the serological tests and cell culture, molecular methods such as PCR are of great importance. Therefore, in this study, our goal was to use a multiple specific PCR assay on L1 and E6 genes of HPV for molecular detection of this virus and its common type's detection in the society. Materials and Methods: After collecting the samples from malignant lesions of various patients, the viral DNA was extracted from paraffin blocks of 50 clinical samples and the PCR method was performed on the mentioned samples by specific primers for L1 and E6 genes together with β-globin (as internal control). The PCR product was analyzed on 2℅ agarose gel and the sensitivity of this test was examined. Results: from among 50 samples of the patients, 33 cases were HPV positive and 17 ones were negative. The sensitivity of this test was 20 copies from recombinant construct containing target genes for each reaction. Conclusion: this study confirmed that the designed PCR with specific primers on L1 and E6 genes of HPV proved to be an accurate method for detecting and determining the HPV types.

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Detection and Typing of Human Papilloma Virus by Multiplex PCR with Type-Specific Primers

Cited by 27 publications

References 26 publications

Relevant Frequency of Multiple Infections with High- and Low-Risk HPV Genotypes among Mexican Women Attending a Tertiary Care Hospital

Relevant Frequency of Multiple Infections with High- and Low-Risk HPV Genotypes among Mexican Women Attending a Tertiary Care Hospital

Simultaneous detection of fungal, bacterial, and viral pathogens in insects by multiplex PCR and capillary electrophoresis

Qualitative-based Multiplex PCR: Diagnosis of Papilloma virus Types 16 & 18 in Samples Taken from Patients with Malignant Lesions

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