Comparison of the detection of Toxocara spp. in the soils of public parks of Ahvaz (southwest of Iran) by PCR and Loop-Mediated Isothermal Amplification (LAMP)

Author(s):  
Forough Kazemi ◽  
Reza Arjmand ◽  
Somayeh Fallahizadeh ◽  
Mehdi Tavalla

Introduction: The infections caused by Toxocara spp. are considered as one of the most important zoonotic diseases in the world, especially in developing countries. Human toxocariasis, particularly in children is acquired by playing in public parks. Hence, the aim of the current study was to detect Toxocara spp. in the soils of public parks of the city of Ahvaz, southwest of Iran, using the PCR and loop-mediated isothermal amplification (LAMP) methods. Methods: In this descriptive cross-sectional study, 260 soil samples were randomly collected from the different public parks of the city of Ahvaz. After preforming zinc sulfate (ZnSO4) flotation technique, the DNA samples were extracted from the isolated Toxocara spp. egg. Lastly, the extracted DNA was used for PCR and LAMP-based molecular detection. Results: Of 260 specimens, 57 (21.9%) samples were found positive for Toxocara spp., using the PCR method, out of that 38 (14.6%) samples were positive for T. canis and 19 (7.3%) samples were positive for T. cati. Also, of 260, 81 (31.1%) cases were positive for Toxocara species, using the LAMP method, among them 51 (19.6%) samples were found positive for T. canis and 30 (11.5%) samples were positive for T. cati. Kappa (κ) coefficient between PCR and LAMP showed a strong agreement (0.766, P-value=0.002). Conclusion: Our data showed a relatively high outbreak of Toxocara spp. in the public parks’ soils of the city, using the PCR and LAMP methods. Since the parasite can cause human toxocariasis particularly in children; thus, the health authorities of the city of Ahvaz and other similar cities, particularly in developing countries must pay more attention to the hygiene of the public parks’ soils.

Author(s):  
Hadi MIRAHMADI ◽  
Raheleh HASANZADEH ◽  
Hamid MALEK RAEESI ◽  
Shirzad FALLAHI ◽  
Mahdi KHOSHSIMA SHAHRAKI ◽  
...  

Background: Toxoplasma gondii (T. gondii) causes an important parasitic infection known as toxoplasmosis, which is a globally distributed important zoonosis. One of the major serious characteristics of T. gondii is its ability to manipulate the behavior of intermediate hosts. We performed a cross-sectional study to determine toxoplasmosis in schizophrenic patients, as one of the major neuropsychiatric disorders, using loop-mediated isothermal amplification (LAMP) technic by targeting parasite B1 gene. Methods: Blood samples were taken from 118 schizophrenic patients hospitalized in tow hospitals including Baharan, Clinic of Psychiatric Ali-ibn-Abi-Talib Hospital (in Zahedan City), and Amir-al Momenin Psychiatric Hospital (in Zabol City), Sistan and Baluchestan Province, southeast Iran in 2016. They were analyzed using LAMP, and compared with the previous data of nested-PCR and serology. Results: Out of the 118 schizophrenic individuals, 56 patients (47.4%) were found to be infected with T. gondii. The diagnosis of toxoplasmosis was confirmed in 41 patients (34.7%) via the nested-PCR. The seroprevalence of toxoplasmosis in schizophrenic patients was 55.9% (66/118). Conclusion: We found a high efficiency of LAMP method in identifying toxoplasmosis and its high prevalence among schizophrenic patients. Our findings could provide viable offer implications for the prevention of schizophrenia.


2021 ◽  
Vol 15 (1) ◽  
pp. e0008996
Author(s):  
Thoko Flav Kapalamula ◽  
Jeewan Thapa ◽  
Mwangala Lonah Akapelwa ◽  
Kyoko Hayashida ◽  
Stephen V. Gordon ◽  
...  

Bovine tuberculosis (TB) caused by Mycobacterium bovis is a significant health threat to cattle and a zoonotic threat for humans in many developing countries. Rapid and accurate detection of M. bovis is fundamental for controlling the disease in animals and humans, and for the proper treatment of patients as one of the first-line anti-TB drug, pyrazinamide, is ineffective against M. bovis. Currently, there are no rapid, simplified and low-cost diagnostic methods that can be easily integrated for use in many developing countries. Here, we report the development of a loop-mediated isothermal amplification (LAMP) assay for specific identification of M. bovis by targeting the region of difference 4 (RD4), a 12.7 kb genomic region that is deleted solely in M. bovis. The assay's specificity was evaluated using 139 isolates comprising 65 M. bovis isolates, 40 M. tuberculosis isolates, seven M. tuberculosis complex reference strains, 22 non-tuberculous mycobacteria and five other bacteria. The established LAMP detected only M. bovis isolates as positive and no false positives were observed using the other mycobacteria and non-mycobacteria tested. Our LAMP assay detected as low as 10 copies of M. bovis genomic DNA within 40 minutes. The procedure of LAMP is simple with an incubation at a constant temperature. Results are observed with the naked eye by a color change, and there is no need for expensive equipment. The established LAMP can be used for the detection of M. bovis infections in cattle and humans in resource-limited areas.


2020 ◽  
Author(s):  
Yuki Higashimoto ◽  
Masaru Ihira ◽  
Yoshiki Kawamura ◽  
Masato Inaba ◽  
Kazuya Shirato ◽  
...  

Coronavirus disease 2019 (COVID-19) has had a major disease burden on many countries around the world. The spread of COVID-19 is anticipated to have a major impact on developing countries including African nations. To establish a point-of-care test for COVID-19, we developed a dry loop mediated isothermal amplification (LAMP) method to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA. We carried out reverse transcription (RT)-LAMP using the Loopamp SARS-CoV-2 Detection kit (Eiken Chemical, Tokyo, Japan). The entire mixture except for the primers is dried and immobilized inside the tube lid. To determine the specificity of the kit, 22 viral genomes associated with respiratory infections, including the SARS coronavirus, were tested. No LAMP product was detected in reactions performed with RNA from these pathogens. The sensitivity of this assay, determined by either a real-time turbidity assay or colorimetric change of the reaction mixture, as evaluated by the naked eye or under illumination with ultraviolet light, was 10 copies/reaction. After the initial validation analysis, we analyzed 24 nasopharyngeal swab specimens collected from patients suspected to have COVID-19. Nineteen (79.2%) of the 24 samples were positive for SARS-CoV-2 RNA, as determined by real-time RT-PCR analysis. Using the Loopamp SARS-CoV-2 Detection kit, we detected SARS-CoV-2 RNA in 15 (62.5%) of the 24 samples. Thus, the sensitivity, specificity, positive predictive value, and negative predictive value of the Loopamp 2019-CoV-2 detection reagent kit were 94.0%, 96.0%, 95.9%, and 94.1%, respectively. The dry LAMP method for detection of SARS-CoV-2 RNA was fast and easy to use, solves the cold chain problem, and therefore represents a promising tool for diagnosis of COVID-19 in developing countries.


2004 ◽  
Vol 28 (6) ◽  
pp. 445-450 ◽  
Author(s):  
Taketoshi Wakabayashi ◽  
Ryoko Yamashita ◽  
Tetsuhiko Kakita ◽  
Mito Kakita ◽  
Tetsuro Oshika

2018 ◽  
Vol 1 (1) ◽  
pp. 12
Author(s):  
Menik Kustriyani ◽  
Ivana Probo Kaeksi ◽  
Tamrin Tamrin

Joint Commission International ( JCI ) required the achievement of 100% five moment hand hygiene for the nurses who have provided care to patients. The adherence of five moments hand hygiene has been done to reduce the incidence of nosocomial infections. The adherence of five moments hand hygiene has been determined by inside and outside factors, and one of the inside factors is the motivation. The research is a qualitative research with cross sectional approach. The number of sample is 153 nurses with the proportionate random sampling technique at the Public Hospital of Loekmono Hadi Kudus. The research instruments used the questionaire and observation sheet. The research showed the result of Rank Spearman test p value = 0,000 with r value = 0.296, positive correlation means that the higher the nurse motivation, the higher the nurse aderence of five moment hand hygiene.


2021 ◽  
Vol 12 (3) ◽  
pp. 389
Author(s):  
Rico Januar Sitorus ◽  
Novrikasari Novrikasari ◽  
Rizma Adliah Syakurah ◽  
Merry Natalia

<p>Antiretroviral treatment side-effects and patient compliance with medical instructions continue to be a growing challenge for HIV/AIDS patients. Arv therapy has resulted in a substantial intervention that has been successful in preventing transmission and opportunistic infection. The main objective of this study was to analyze the association between side-effects of ARV therapy and medication adherence as well as another potential confounding such as opportunistic infection, family support, stress level, knowledge of ARV, marital status, and occupation. This study is a quantitative approach by using cross-sectional methods. A total of 244 respondents from 1.180 patients with confirmed HIV registered in the Care Support and Treatment (CST) service and Sriwijaya Community in the City of Palembang to respond to the survey. Non-random sampling was used to collect the samples. As the result, the majority of the respondents were male (84,43 %), ≥30 years old (57,4%), and secondary school graduates (52%). After adjusting with stress and opportunistic infection variable, a Multivariate regression analysis revealed a positive relationship between side effects and medication adherence (p-value of 0,041; 0,05), OR Adj 2,131 (1,190-4,988). PLHV who had adverse effects had a 2.131 times worse adherence rate than those who did not. In conclusion, medication adherence must be greatly improved in light of therapeutic side effects, stress levels, and opportunistic infection.</p>


2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Zhang Tie ◽  
Wang Chunguang ◽  
Wei Xiaoyuan ◽  
Zhao Xinghua ◽  
Zhong Xiuhui

To develop a rapid detection method ofStaphylococcus aureususing loop-mediated isothermal amplification (LAMP), four specific primers were designed according to six distinct sequences of thenucgene. In addition, the specificity and sensitivity of LAMP were verified and compared with those of PCR. Results showed that the LAMP reaction was completed within 45 min at 62.5°C, and ladder bands were appeared in LAMP products analyzed by gel electrophoresis. After adding 1x SYBR Green l, the positive reaction tube showed green color and the negative reaction tube remained orange, indicating that the LAMP has high specificity. The minimal detectable concentration of LAMP was1×102 CFU/mL and that of PCR was1×104 CFU/mL, indicating that the LAMP was 100 times more sensitive than the PCR. The LAMP method for detection ofStaphylococcus aureushas many advantages, such as simple operation, high sensitivity, high specificity, and rapid analysis. Therefore, this method is more suitable for the rapid on-site detection ofStaphylococcus aureus.


2011 ◽  
Vol 47 (No. 4) ◽  
pp. 140-148 ◽  
Author(s):  
N. Rostamkhani ◽  
A. Haghnazari ◽  
M. Tohidfar ◽  
A. Moradi

In an attempt to speed up the process of screening of transgenic cotton (G. hirsutum L.) plants, a visual and rapid loop-mediated isothermal amplification (LAMP) assay was adopted. Genomic DNA was extracted from fresh leaf tissues of T<sub>2</sub> transgenic cotton containing chitinase (chi) and cry1A(b) genes. Detection of genes of interest was performed by polymerase chain reaction (PCR), LAMP and real-time PCR methods. In LAMP assay the amplification was performed after 30 min at 65&deg;C when loop primers were involved in the reaction. The involvement of loop primers decreased the time needed for amplification. By testing serial tenfold dilutions (10<sup>&ndash;1</sup> to 10<sup>&ndash;8</sup>) of the genes of interest, the detection sensitivity of LAMP was found to be 100-fold higher than that of PCR. The rapid DNA extraction method and LAMP assay can be performed within 30 min and the derived LAMP products can be directly observed as visually detectable based on turbidity in the reaction tube. The accuracy of LAMP method in the screening of transgenes was confirmed by PCR and real-time PCR. The developed method was efficient, rapid and sensitive in the screening of cotton transgenic plants. This method can be applied to any other crops.


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