Molecular Methods in Immunohematology: Use of the PCR-SSP Technique

Abstract Background Canine vector-borne diseases (CVBDs) are caused by a wide range of pathogens transmitted by arthropods. They have been an issue of growing importance in recent years; however, there is limited information about the vector-borne pathogens circulating in Portugal. The aim of the present study was to detect canine vector-borne bacteria and protozoa of veterinary and zoonotic importance using molecular methods. Methods One hundred and forty-two dogs from Lisbon, southern Portugal, were tested: 48 dogs from a veterinary hospital clinically suspected of vector-borne diseases and 94 apparently healthy dogs from shelters. Anaplasma spp./Ehrlichia spp., Babesia/Theileria spp., Hepatozoon spp., and Mycoplasma spp. infections were detected by PCR from blood samples and examined under light microscopy. Other information including clinical status and diagnostic test results were collected for each animal. Results Infections were detected by PCR in 48 (33.80%) dogs. Single infections were found in 35 dogs (24.64%), and co-infections were found in 13 (9.15%) dogs. Twenty-nine (20.42%) dogs were positive for Hepatozoon spp., 15 (10.56%) for Mycoplasma spp., 11 (7.75%) for Anaplasma spp./Ehrlichia spp., and six (4.21%) for Babesia spp. DNA sequencing was used to identify Babesia vogeli (2.81%), Babesia canis (1.40%), Hepatozoon canis (20.42%), Mycoplasma haematoparvum (2.11%), Mycoplasma haemocanis (8.45%), Anaplasma platys (7.04%), and Ehrlichia canis (0.70%). Conclusions This is the first molecular identification of B. canis and M. haematoparvum in dogs from southern Portugal. This study highlights the importance of molecular methods to identify CVBD pathogens in endemic areas and helps to guide the clinical approach of veterinarians in practice.

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Classical and molecular methods in differentiation of mantle cell lymphoma and small lymphocytic lymphoma in composite lymphoma: a case report

Journal of Hematopathology ◽

10.1007/s12308-020-00426-4 ◽

2021 ◽

Author(s):

Michał Szymczyk ◽

Grzegorz Rymkiewicz ◽

Zbigniew Bystydzieński ◽

Małgorzata Szostakowska-Rodzoś ◽

Renata Zub ◽

...

Keyword(s):

Case Report ◽

Mantle Cell Lymphoma ◽

Cell Lymphoma ◽

Molecular Methods ◽

Small Lymphocytic Lymphoma ◽

Composite Lymphoma ◽

Mantle Cell

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Pediatric Invasive Pneumococcal Disease Three Years after PCV13 Introduction in the National Immunization Plan—The Continued Importance of Serotype 3

Microorganisms ◽

10.3390/microorganisms9071428 ◽

2021 ◽

Vol 9 (7) ◽

pp. 1428

Author(s):

Catarina Silva-Costa ◽

Joana Gomes-Silva ◽

Lúcia Prados ◽

Mário Ramirez ◽

José Melo-Cristino ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Antimicrobial Resistance ◽

Conjugate Vaccine ◽

Invasive Pneumococcal Disease ◽

Pneumococcal Disease ◽

Molecular Methods ◽

Vaccine Uptake ◽

Serotype Distribution ◽

Conjugate Vaccines ◽

National Immunization

The introduction of pneumococcal conjugate vaccines PCV7 and PCV13 led to decreases in incidence of pediatric invasive pneumococcal disease (pIPD) and changes in serotype distribution. We evaluated the consequences of higher vaccine uptake after the introduction of PCV13 in the National Immunization Plan (NIP) in 2015. Besides culture and conventional serotyping, the use of molecular methods to detect and serotype pneumococci in both pleural and cerebrospinal fluid samples contributed to 30% of all pIPD (n = 232) in 2015–2018. The most frequently detected serotypes were: 3 (n = 59, 26%), 10A (n = 17, 8%), 8 (n = 16, 7%) and 19A (n = 10, 4%). PCV13 serotypes still accounted for 46% of pIPD cases. Serotypes not included in any currently available conjugate vaccine (NVT) are becoming important causes of pIPD, with the increases in serotypes 8 and 33F being of particular concern given the importance of serotype 8 in adult IPD and the antimicrobial resistance of serotype 33F isolates. This study highlights the importance of using molecular methods in pIPD surveillance since these allowed a better case ascertainment and the identification of serotype 3 as the leading cause of pIPD. Even in a situation of vaccine uptake >95% for 3 years, PCV13 serotypes remain important causes of pIPD.

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